Cytogenetics

BACHELOR OF SCIENCE IN MEDICAL TECHNOLOGY
NATIONAL UNIVERSITY – MALL OF ASIA

CYTOGENETICS
BSMT II – MED212 || MS. KYLE MIRAH B. SUMAYAO, RMT, MSMLS
THE GENETIC MATERIAL o Thus, DNA is the genetic material

DNA Is the Genetic Material Discovering the Structure of DNA
Francis Crick 1953 Phoebus Levine
"A genetic material must carry out two jobs: • Russian-American biochemist
duplicate itself and control the development of • Identified the 5-carbon sugars ribose in
the rest of the cell in a specific way.” 1909 and deoxyribose in 1929
History of DNA • Revealed chemical distinction between
Friedrich Miescher, 1871 RNA and DNA
• Swiss physician and biochemist o RNA has ribose
• Isolated nuclei from white blood cells in pus o DNA has deoxyribose
• Found an acid substance with nitrogen and • Discovered that the three parts of a
phosphorus nucleotide are found in equal proportions:
o He called it nuclein o Sugar
o Later, it was called nucleic acid o Phosphate
Archibald Garrod, 1902 o Base
• English physician • Deduced that a nucleic acid building block
• Linked inheritance of inborn errors of must contain one of each component
metabolism with the lack of particular Erwin Chargaff, 1951
enzymes • Austrian-American biochemist
Frederick Griffith, 1928 • Analyzed base composition of DNA from
• English microbiologist various species and observed regular
• Worked with Streptococcus pneumoniae relationships:
bacteria, which exists in two types: o Adenine (A) + Guanine (G) = Thymine
o Type S (Smooth) = Enclosed in a (T) + Cytosine (C)
polysaccharide capsule o A = T and C = G
o Type R (Rough) = No capsule Rosalind Franklin and Maurice Wilkins,
• Termed the conversion of one bacterial type 1952
into another as transformation • English scientists
Avery, MacLeod, and McCarty, 1944 • Used a technique called X-ray diffraction
• American physicians o Deduced the overall structure of the
• Treated lysed S bacteria with protease and molecule from the patterns in which the X
DNase rays were deflected
• Only DNase prevented transformation • Distinguished two forms of DNA
• Thus, DNA is the transforming principle o “A” form, which is dry and crystalline
o Can convert type R bacteria into S o “B” form, which is wet and cellular
Alfred Hershey and Martha Chase, 1953 • It took Franklin 100 hours to obtain “photo
American microbiologists 51” of the B-form of DNA
• Used E.coli bacteria infected with a virus that • Franklin reasoned that the DNA is a helix
consisted of a protein head surrounding DNA with symmetrically organized subunits.
• Grew a batch of virus in a medium containing
35S and 32P
• Blender experiments showed that the virus
transfers DNA, not protein, into a bacterial
cell
P a g e 1 | Diamola, Nikka V.
CYTOGENETICS
James Watson and Francis Crick o A phosphate group

• Did not perform any experiments o A nitrogenous base; one of four types:
o Rather, they used the earlier research o Adenine (A), Guanine (G) =
and inferences from model building with Purines
cardboard cutouts to solve the structure o Cytosine (C), Thymine (T) =
of DNA Pyrimidines
The Road to the Double Helix
Investigator Contribution Timeline

Isolated nuclein in
Friedrich Miescher white blood cell 1869
nuclei
Transferred killing
Frederick Griffith ability between types 1928
of bacteria
Oswald Avery,
Discovered that DNA
Colin
transmits killing 1940s
MacLeod, and
ability in bacteria
Maclyn McCarty
Determined that the
part of a virus
Alfred Hershey that infects and
1950
and Martha Chase replicates is its
nucleic acid and not
its protein
Phoebus Levene,
Discovered DNA
Erwin Chargaff,
components, 1909-
Maurice
proportions, and early 1950s
Wilkins, and
positons
Rosalind Franklin
Elucidated DNA’s
James Watson
three- 1953
and Francis Crick
dimensional structure
Had his genome
James Watson 2008 • Nucleotides are joined into chains.
sequenced
o Phosphodiester bonds form between the
deoxyribose sugars and the phosphates.
DNA Structure
o This creates a continuous sugar-
• Gene is a section of a DNA molecule phosphate backbone.
o Sequence of building blocks specifies the
• DNA Consists of Two Chains of Nucleotides
sequence of amino acids in a particular in an Antiparallel Configuration
protein.
• Two polynucleotide chains align forming a
• A single building block is a nucleotide.
double helix.
o Each nucleotide is composed of:
o A deoxyribose sugar
CYTOGENETICS
o The opposing orientation (head-to-toe) is • The key to the constant width of the double
called antiparallelism. helix is the specific pairing of purines and
pyrimidines via hydrogen bonds
• The complementary base pairs are:
o Adenine and guanine
o Cytosine and thymine
• Hydrogen bonds hold the base pairs together
DNA Is Directional
• Note that one strand of the doublehelix runs
in a 5’ to 3’ direction, and the other strand
runs in a 3’ to 5’ direction.
DNA Is Highly Condensed
• Scaffold proteins form frameworks that guide
DNA strands.
• Antiparallel nature of the DNA double helix • The DNA coils around proteins called
becomes apparent when the carbons in the histones, forming a bead-on-a-string-like
sugar are numbered. structure.
o Carbons are numbered from 1 to 5. o The bead part is called the nucleosome.
• DNA wraps at several levels, until it is
compacted into a chromatid.
• Chromosome substance is called chromatin.
CYTOGENETICS
• Nontemplate strand of the DNA double helix

is called the coding strand.
Transcription
Nucleic Acids
• There are two types of nucleic acids:
o RNA
o DNA
• Both consist of sequences of nitrogen-
containing bases joined by sugar-phosphate
• When chromatin is loose (not condensed into backbones.
chromosomes that are visible upon staining), o However, they differ in several aspects.
it forms loops at about 10,000 places in the How DNA and RNA Differ
genome.
• An “anchor” protein called CTCF brings
together parts of the DNA sequence within DNA RNA
the same long DNA molecule to form the
1. Usually single-
overall “loop-ome” structure. 1. Usually double- stranded
stranded
2. Uracil as a base
2. Thymine as a base
3. Deoxyribose as the 3. Ribose as the sugar

sugar
4. Carries protein-
encoding information
4. Maintains protein- and controls
encoding information how information is
used
5. Can function as an
5. Cannot function as an enzyme
enzyme
RNA Structure and Types
6. Short-lived
• RNA is the bridge between gene and protein. 6. Persists
• Bases of an RNA sequence are
complementary to those of one strand of the DNA and RNA Differences
double helix, called the template strand. • DNA
• RNA polymerase builds an RNA molecule.
CYTOGENETICS
o Stores RNA- and protein-encoding Associates

information, and transfers information to with proteins
daughter cells to form
• RNA ribosomes,
o Carries protein-encoding information, and Ribosomal RNA (rR which structur
100 to 3,000
helps to make proteins NA) ally
support and
catalyze
protein
synthesis
Transports
specific
amino acids
Transfer
75 to 80 to the
RNA (tRNA)
ribosome for
protein
synthesis
mRNA
• Carries information that specifies a particular
protein
• Three mRNA bases in a row form a codon
which specifies a particular amino acid
• Most mRNAs are 500–4500 bases long
• Differentiated cells produce certain mRNA
molecules called transcripts
Types of RNA o Information in the transcripts is used to
• There are three major types of RNA: manufacture the encoded proteins
o Messenger RNA or mRNA rRNA
o Ribosomal RNA or rRNA • Most rRNAs are from 100–3000 nucleotides
o Transfer RNA or tRNA long
• Other classes of RNA control gene • Associate with proteins to form ribosomes
expression • Ribosomes consist of two subunits that join
Major Types of RNA during protein synthesis
• rRNAs provide structural support
Size o Some are catalysts (ribozymes) and
(number others help align the ribosome and
Type of RNA Function
of nucleotid mRNA
es)
Encodes
Messenger RNA (m 500 to
amino acid
RNA) 4,500+
sequence
CYTOGENETICS
tRNA
• Binds an mRNA codon and a specific amino
acid
• Only 75–80 nucleotides long
o The 2-D shape is a cloverleaf shape
o The 3-D shape is an inverted L
• Has two ends:
o The anticodon is complementary to an
o mRNA codon
o The opposite end strongly bonds to a
specific amino acid
CYTOGENETICS
DNA REPLICATION AND PROTEIN o Two identical nucleotide chains are built
SYNTHESIS from one, as the bases form pairs.
DNA Replication • A site where DNA is locally opened is called
• DNA replication is semiconservative. a replication fork.
o Two identical double helices are formed Overview of DNA Replication
from one original, parental double helix. 1. Parent DNA molecule.
o Each new DNA double helix 2. Parental strands unwind and separate at
conserves half of the original. several points.
Matthew Meselson and Franklin Stahl, 1957 3. Each parental strand provides a template for
• Demonstrated the semiconservative DNA polymerase to bind complementary
mechanism of DNA replication with a series bases, A with T and G with C.
of density shift experiments 4. Sugar-phosphate backbones of daughter
• Labeled replicating DNA from bacteria with a strands close.
heavy form of nitrogen and traced its pattern
of distribution
o Higher-density nitrogen was incorporated
into one strand of each daughter double
helix
DNA Replication Is Semiconservative
Enzymes in DNA Replication
Steps of DNA Replication

• DNA replication occurs during the S phase of
the cell cycle, prior to cell division. • Helicase unwinds parental double helix.
• When DNA is replicated, it separates and • Binding proteins stabilize separate strands.
hydrogen bonds holding the base pairs • Primase adds short primer to template
together break. strand.
CYTOGENETICS
• DNA polymerase binds nucleotides to form

new strands.
• Ligase joins Okazaki fragments and seals
other nicks in sugar-phosphate backbone.
Activities at the Replication Fork
8. Enzymes remove RNA primers. Ligase seals

sugar-phosphate backbone.
Replication Bubbles
• The sites of replication resemble bubbles that
coalesce as the daughter double helices
form.
1. Helicase binds to origin and separates
strands.
2. Binding proteins keep strands apart.
3. Primase makes a short stretch of RNA on the
DNA template.
4. DNA polymerase adds DNA nucleotides to

the RNA primer.
5. DNA polymerase proofreading activity Polymerase Chain Reaction (PCR)
checks and replaces incorrect bases. • DNA amplification technique
o Uses DNA polymerase to rapidly replicate
a specific DNA sequence in a test tube
• Yields more than 10 billion copies of the
target DNA sequence
• Useful in forensic investigations to amplify
small DNA samples
Steps in Amplifying DNA Using PCR
6. Continuous strand synthesis continues in a 5’

to 3’ direction.
7. Discontinuous synthesis produces Okazaki
fragments on the 5’ to 3’ template.
CYTOGENETICS
1. Select target sequence in virus genome.

2. Primers.
• Preparation
3. Free nucleotides
4. Heat-resistant polymerase
• Temperature shift
5. Target sequence (Up)
6. Heat separates strands. (Down)
• Hybridization
7. Primers hybridize due to base
complementarity.
8. DNA fills in.
9. Repeat process many times.
Gene Expression
• Amplification
• DNA of the human genome which encodes
protein is called the exome.
o However, this represents only a small part
of the genome.
• Much of the human genome controls protein
synthesis.
o Including the time, speed, and location
• Genes encode 20,325 types of proteins.
• Production of protein from instructions on the
Proteins DNA
• The 20,325 or so proteins the human has are • Gene expression requires several steps:
coded by protein coding genes, termed the o Transcription = Synthesizes an RNA
exome. molecule
• Proteins serve many vital functions; some of o Translation = Uses the information in the
these functions are contractile, regulatory, RNA to manufacture a protein by aligning
enzymatic, structural, transport, immunity, and joining specified amino acids
and clotting. • Folding of the protein into specific 3-D form
Central Dogma
• Proteins are comprised of one or more long
chains of amino acids called polypeptides. • The directional flow of genetic information
• There 20 naturally occurring amino acids.
• Each amino acid has a central carbon atom
that bonds to an amino group (NH2) and an
acidic group (COOH), a hydrogen atom (H),
and a variable “R” group.
• It is the R group that distinguishes the 20
types of amino acids.
Amino Acid Structure
CYTOGENETICS
Transcription Setting the Stage for Transcription to

Begin
Transcription Factors
• Interact and form an apparatus that binds
DNA at certain sequences
• Initiates transcription at specific sites on
chromosomes
• Respond to signals from outside the cell
• Link the genome to the environment
• Mutations in transcription factors may cause
a wide range of effects
Steps of Transcription
• Transcription is described in three steps:
o Initiation Transcription of RNA from DNA
o Elongation
o Termination
• In transcription initiation, transcription factors
and RNA polymerase are attracted to a
promoter.
• RNA polymerase joins the complex, binding
in front of the start of the gene sequence.
• In transcription elongation, enzymes unwind
the DNA double helix.
o Free RNA nucleotides bond with exposed
complementary bases on the DNA
template strand.
o RNA polymerase adds the RNA
nucleotides, in the sequence the DNA
specifies.
• A terminator sequence in the DNA indicates
where the gene’s RNA-encoding region
ends.
Simultaneous Transcription of mRNAs

• Several mRNAs may be transcribed from the
same template DNA strand at a time.
CYTOGENETICS
Translation
• Assembles a protein using the information in
the mRNA sequence
o Particular mRNA codons correspond to
particular amino acids
• Occurs on the ribosome
RNA Processing
• In eukaryotes, mRNA must exit the nucleus
to enter the cytoplasm.
• Several steps process pre-mRNA into mature
mRNA.
o A methylated cap is added to the 5’ end. The Genetic Code
o Recognition site for protein synthesis • The correspondence between the chemical
• A poly A tail is added to the 3’ end. languages of mRNA and proteins
o Necessary for protein synthesis to begin • In the 1960s, researchers used logic and
and stabilizes the mRNA clever experiments on simple genetic
• Splicing occurs. systems to decipher the genetic code
o Introns (“intervening sequences”) are
removed.
o Ends of the remaining molecule are
spliced together.
o Exons are parts of mRNA that remain,
translated into amino acid sequences.
o Note that introns may outnumber and
outsize exons.
• mRNA is proofread and the mature mRNA is
sent out of the nucleus.
Alternate Splicing
• Mechanism of combining exons of a gene in
different ways:
o Cell types can use versions of the same
protein in slightly different ways in
different tissues
Messenger RNA Processing – The
Maturing of the Message
CYTOGENETICS
o Different codons that specify the same

• It is a triplet code. amino acid are termed synonymous
o Three successive mRNA bases form a codons
codon. o Nonsynonymous codons encode
o There are 64 codons. different amino acids
o Altering the DNA sequence by one or two Three at a Time
bases produced a different amino acid
sequence due to disruption in the reading
frame.
o Adding a base at one point and
deleting a base at another point
disrupted the reading frame between
the sites.
• It is nonoverlapping.
o In an overlapping DNA sequence, certain
amino acids would follow others,
constraining protein structure
• It includes controls.
o Includes directions for starting and
stopping translation
o An open reading frame does not
include a stop codon
• It is universal.
o Evidence that all life evolved from a
common ancestor
CYTOGENETICS
Reading Frame • GGA bonds to its complementary anticodon,

which is part of a free tRNA that carries the
amino acid glycine.
o Two amino acids attached to their tRNAs
align.
• Positions of the sites on the ribosome remain
the same, cover different parts of the mRNA
as the ribosome moves.
o The P site bears growing amino acid
chain.
o The A site holds the next amino acid to be
added to the chain.
• Amino acids link by a peptide bond, with the
Translation – Building a Protein help of rRNA that functions as a ribozyme.
• Requires mRNA, tRNAs with amino acids, • The polypeptide builds one amino acid at a
ribosomes, energy molecules (ATP, GTP) time.
and protein factors o Each piece is brought in by a tRNA
• Divided into three steps: whose anticodon corresponds to a
o Initiation consecutive mRNA codon as the
o Elongation ribosome moves down the mRNA.
o Termination Building a Polypeptide
Translation Initiation
• The leader sequence of the mRNA forms H
bonds with the small ribosomal subunit.
• The start codon (AUG) attracts an initiator
tRNA that carries methionine.
• This completes the initiation complex.
Translation Begins as the Initiation
Complex Forms
Translation Termination
• Occurs when a stop codon enters the A site
Translation Elongation of the ribosome
• The large ribosomal subunit joins. • A protein release factor frees the polypeptide
CYTOGENETICS
• The ribosomal subunits separate and are o Secondary (2°) structure

recycled o Tertiary (3°) structure
• New polypeptide is released o Quaternary (4°) structure
Termination Translation Four Levels of Protein Structure
1. Primary structure – The sequence of amino

Multiple Copies of a Protein Can Be Made acids in a polypeptide chain
Simultaneously 2. Secondary structure – Loops, coils, sheets,
• The closer to the end of the gene, the longer or other shapes formed by hydrogen bonds
the polypeptide between neighboring carboxyl and amino
groups
3. Tertiary structure – Three dimensional
forms shaped by bonds between R groups,
interaction between R groups and water
4. Quaternary structure – Protein complexes
formed by bonds between separate
polypeptides
Protein Folding
• Proteins begin to fold after the amino acid
Protein Structure chain winds away from the ribosome.
• Proteins fold into one or more 3-D shapes or o First few amino acids in a protein
conformations secreted in a membrane form a “signal
o Based on attraction and repulsion sequence.”
between atoms of proteins, and o Leads it and the ribosome into a pore
interactions of proteins with chemicals in in the ER membrane
the environment o Not found on proteins synthesized on
• There are four levels for protein structure: free ribosomes
o Primary (1°) structure • Chaperone proteins
CYTOGENETICS
o Stabilize partially folded regions in their Diseases Associated with Protein

correct form Misfolding
o Prevent a protein from getting stuck in an
intermediate form
Disease Misfolded Protein(s)
o Developed into drugs to treat diseases
that result from misfolded proteins Amyloid
Protein Misfolding Alzheimer disease beta precursor
• Misfolded proteins are tagged with ubiquitin. protein, tau proteins
• Protein with more than one tag is taken to a
proteasome, a tunnel-like multiprotein Familial amyotrophic Superoxide
structure. lateral sclerosis dismutase, TDP-43
o As the protein moves through the tunnel, Tau proteins, TDP-
it is straightened and dismantled. Frontotemporal dementia
43
o Proteasomes also destroy properly folded
proteins that are in excess or no longer Huntington disease Huntingtin
needed.
Parkinson disease Alpha synuclein
Lewy body dementia Alpha synuclein

Phenylalanine
PKU
hydroxylase
Prion diseases Prion protein

• All but Huntington disease are genetically
heterogeneic; that is, abnormalities in
different proteins cause similar syndromes.
Prion Diseases
• Proteins misfold from a mutation, or by
having more than one conformation. • They have been found in 85 species
o A mutation alters the attractions and o Scrapie in sheep
repulsions between parts of the protein. o In humans
o Kuru
o Prion protein can fold into any of several
o Creutzfeldt-Jakob disease
conformations.
• Disease
o Moreover, it can be passed on to other
o Creutzfeldt-Jakob disease
proteins upon contact, propagating
o Fatal familial insomnia
like an infectious agent.
o Gerstmann-Straüssler-Scheinker disease
• In several disorders that affect the brain, the Prions
misfolded proteins aggregate.
o The protein masses that form clog the
proteasomes and inhibit their function.
• Different proteins are affected in different
disorders.
CYTOGENETICS
GENE MUTATION o A person with a somatic mutation has

The Nature of Gene Variants somatic mosaicism
• The terms mutation and polymorphism each Somatic Mosaicism
denote a genetic change from the wild type • When somatic cells mutate in an individual,
(most common form). not all the cells in that individual have that
• A mutation is change in a DNA sequence is mutation.
rare in a population and typically affects the • Because not all the cells are affected by the
phenotype. mutation, the individual has somatic
• A polymorphism simply means many forms. mosaicism.
It is a change in a gene that is less rare than
a mutation and more prevalent in a
population.
• The sequencing of many genomes shows
that a DNA base change that is a mutation in
one population may be a harmless
polymorphism in another, due to the effects
of other genes and different environments.
• The distinction between the two terms
reflects gene function as well as frequency.
• Because of the confusion between the terms
mutation and polymorphism, the term “gene
variant” is being used for both terms.
• The effect of mutations vary.
o “Loss-of-function” mutations—Recessive
o “Gain-of-function” mutations—Dominant
• The term mutant refers to phenotype. Mutations Alter Proteins
o Usually connotes an abnormal or • Identifying how a mutation causes symptoms
unusual, or even uncommon variant that has clinical applications
is nevertheless “normal” • Examples of mutations that cause disease:
• Mutations are important to evolution. o Beta globin gene
• Our evolutionary relatedness to other species o Collagen genes
allows us to study many mutations in Collagen Disorders
nonhuman species.
Germline and Somatic Mutation Signs and
Mutations
Disorder Symptoms
• Germline mutation (Genotype)
(Phenotype)
o Change occurs during the DNA
Mutations in any
replication that precedes meiosis
of three genes
o Transmitted to the next generation of
(COL4A3,
individuals COL4A4, COL4A
• Somatic mutation Alport Deafness and
5) affect type IV
o Happens during DNA replication before a syndrome inflamed kidneys
collagen, which
mitotic cell division disrupts
o Affect only cells that descend from tissue boundarie
changed cell s.
CYTOGENETICS
Deletion, of
insertion, or collagen molecul
missense es.
mutation
Chondrodys replaces Gly wit Stunted growth, How Mutations Cause Disease
plasia h bulky amino deformed joints
acids
in COL2A1 type Mutatio Signs and
II ns Symptoms
Disease Protein
collagen gene. (Genoty (Phenotyp
pe) e)
Mutation
in COL7A1 gene Missing
that encodes amino
type acid or
Dystropic other
VII collagen Skin blisters
epidermoly variant al
breaks down upon any touch
sis bullosa ters
fibrils that
attach conforma
epidermis tion of
to dermis. chloride
Cystic channels Frequent
Diverse
fibrosis trans in certain lung infectio
mutations in at Cystic fibr
membrane re epithelial n, pancreati
least a dozen osis
Ehlers- Stretchy, gulator (CFT cell plas c insufficien
genes
Danlos easily scarred R) ma cy
affect collagens
syndrome skin, lax joints membra
or the molecules
nes.
to which they
Water
bind.
enters ce
Missense lls,
mutation in α1 drying
Osteoarthrit collagen gene out
Painful joints
is (COL1A1) substi secretion
tutes Cys for s.
Arg.
Deletion
Inactivation eliminate
of α1 collagen s dystrop
Osteogenes gene hin, whic
Easily broken
is (COL1A1 or COL h
bones; blue eye
imperfecta 1A2) reduces normally
whites; deafness
type I number of Duchenne binds Gradual loss
collagen triple muscular inner of
helices by 50% Dystrophin
dystroph face muscle func
Nonsense y of muscl tion
mutations e cell to
Joint
in type II plasma
Stickler pain, degenerati
procollagen membra
syndrome on of vitreous
gene (COL2A1 o ne. Muscl
gel and retina
r COL11A1) es
reduce number weaken.
CYTOGENETICS
Deficient cause
LDL aneurys
receptors m (bursti
Familial h High blood
cause ch ng).
yperchole cholesterol,
LDL receptor olesterol
sterolemi early heart Defect in
to
a disease protein
accumula
that nor
te in
blood. mally sup
Pigmented
presses
Absent skin patches
activity
or Neurofib and
Neurofibro of a
deficient romatosi benign tumors
Slow or min gene that
clotting f s type 1 of
Hemophili absent causes
Factor IX actor cau nervous tissue
aB blood clottin cell
ses hard- beneath skin
g division; l
to- eading to
control abnormal
bleeding. growths.
Allelic Diseases
Mutatio
Signs and
ns
Disease Protein Symptoms Ge
(Genoty Function Associated Diseases
(Phenotype) ne
pe)
Extra Menkes (“kinky
ATP Copper
bases hair”) disease;
7A transport
add peripheral neuropathy
amino aci DM Dystrophin m Duchenne and Becker
ds to the D uscle protein muscular dystrophy
protein, Encodes
Uncontrollable
Huntingt which fibrillin-
movements, p
on disea Huntingtin impairs c 1, which for
ersonality
se ertain ms tiny Marfan syndrome; systemic
changes FBN
transcript fibrils sclerosis (scleroderma;
ion 1
outside cells; “stiff skin syndrome”)
factors a a connective
nd tissue protei
proteaso n
mes. FGF Fibroblast gr
Deficient Two types of dwarfism
R3 owth factor
proteins Long Glucocerebro Gaucher disease; Parkinson
in limbs, weaken GBA
Fibrillin or t sidase disease
lenses ca ed aorta,
ransformin Presenilin 1
Marfan s use spindly
g growth (enzyme
yndrome cataracts fingers, sunke
factor β rec PSE part that Acne inversa; Alzheimer
and in n chest,
eptor N1 trims disease
the lens dislocatio
membrane p
wall of n roteins)
the aorta
CYTOGENETICS
Oncogene Multiple • Caused by mutagens, many are also

RET (causes endocrine neoplasia; Hirsch carcinogens and cause cancer
cancer) sprung disease • Examples:
TRP Calcium chan Peripheral neuropathy; o Alkylating agents—Remove a base
V4 nel spinal muscular atrophy o Acridine dyes—Add or remove base
o X rays—Break chromosomes
Causes of Mutations o UV radiation—Creates thymine dimers
• Mutations may occur spontaneously or by • Site-directed mutagenesis—Changes a gene
exposure to a chemical or radiation. in a desired way
• An agent that causes a mutation is called a Exposure to Mutagens
mutagen. • Some mutagen exposure is unintentional
Spontaneous Mutation o Workplace
• De novo or new mutations o Industrial accidents
• Not caused by exposure to known mutagen o Chernobyl
• Result from errors in DNA replication • Medical treatments
• DNA bases have slight chemical instability • Weapons
• Exist in alternating forms called tautomers • Natural sources
• As replication fork encounters unstable o Cosmic rays, sunlight, earth’s crust
tautomers, mispairing can occur Types of Mutations
• Mutations can be classified in several ways
o By whether they remove, alter, or add a
function
o By exactly how they structurally alter DNA
• A sentence comprised of three-letter words
analogies to the effects of mutations on a
gene’s DNA sequence:
Spontaneous Mutation Rate

• Rate differs between genes. Point Mutations
o Larger genes usually have higher • A change of a single nucleotide
mutation rates. • Transition = Purine replaces purine or
• Each individual has multiple new mutations. pyrimidine replaces pyrimidine
• Mitochondrial genes mutate at a higher rate o A to G or G to A or
than nuclear genes because they cannot o C to T or T to C
repair their DNA.
Induced Mutations
CYTOGENETICS
• Transversion = Purine replaces pyrimidine Single-base

or pyrimidine replaces purine deletion in
o A or G to T or C alpha globin
o T or C to A or G Hemoglobin gene causes fra
Consequences of Point Mutations Clinically silent (Hb) Wayn meshift,
• Missense mutation = Replaces one amino e changing amino
acids 139-141
acid with another
and adding
• Nonsense mutation = Changes a codon for amino acids
an amino acid into a stop codon Nine extra
o Creates truncated proteins that are often bases add
nonfunctional three amino
• A stop codon that is changed to a coding Hb Grady acids between
codon lengthens the protein amino acids
Splice Site Mutations 118 and 119 of
• Alters a site where an intron is normally alpha chain
removed from mRNA Change from
arginine
• Can affect the phenotype if: Hb Chesap
Oxygen binding to leucine at
o Intron is translated or exon skipped eake
amino acid 92
o Example: CF mutation of beta chain
• Exon is skipped Change from
o Example: Familial dysautonomia (FD) tyrosine to
Deletions and Insertions Hb McKees STOP codon
• The genetic code is read in triplets Rocks at amino acid
• Nucleotides changes not in multiples of 3 145 in beta
lead to disruptions of the reading frame chain
• Cause a frameshift mutation and alter amino Change from
STOP codon to
acids after mutation Hb Constan
Anemia glutamine elong
• Nucleotide changes in multiples of 3 will not t Spring
ates alpha
cause a frameshift chain
o But they can still alter the phenotype Change from
• A deletion removes genetic material glutamic
o Male infertility: Tiny deletions in the Y acid to valine at
Hb S
• An insertion adds genetic material amino acid 6 in
o Gaucher disease: Insertion of one base beta chain
causes sickling
• A tandem duplication is an insertion of
identical sequences side by side Amino acid 6
Hb Leiden deleted from
o Charcot-Marie-Tooth disease: Tandem
beta chain
duplication of 1.5 million bases
Change from
Globin Mutations
glutamic acid to
Protection agains lysine at amino
Associated Phe Hb C
Name Mutation t malaria acid 6 in beta
notype chain causes
sickling
CYTOGENETICS
Not All Mutations Impact Protein Function

• Silent mutations are mutations that do not
alter the encoded amino acid
• Example:
o A mutation from CAA to CAG alters the
DNA, but the protein sequence remains
unchanged
o CAA and CAG both code for glutamine
o These are called synonymous codons
• A missense mutation alters the encoded
amino acid to another amino acid.
o Creates a nonsynonymous codon
• Some nonsynonymous mutations are
conservative; Encode a chemically similar
amino acid and may not alter function.
• The impact of a missense mutation is not
predictable from protein sequence alone.
• A conditional mutation produces a
phenotype under particular conditions or
environments.
• Glucose 6-phosphate dehydrogenase
enzyme responds to oxidants, chemicals that
strip electrons from other molecules.
• High levels of oxidants occur when eating
fava beans or taking certain antimalarial
drugs.
CYTOGENETICS
DNA TECHNOLOGIES Attempts to enforce patents on non-

Patenting DNA protein-encoding parts of the
• Biotechnology is the use or alteration of 2003 human genome anger researchers
cells or biological molecules for specific who support open access to the
applications. information.
• A transgenic organism has DNA from Patent requirements must embrace a
2007 new, more complex definition of a
different species.
gene.
• Recombinant DNA comes from more than
Patents on breast cancer genes
one type of organism. 2009
challenged.
• Both are possible because of the universality Direct-to-consumer genetic testing
of the genetic code. companies struggle to license DNA
• 2010
Mice containing the jellyfish gene for green patents for multigene and SNP
fluorescent protein (GFP) association tests.
Patents on breast cancer genes
invalidated.
U.S. government considers changes to
2011
gene patent laws.
U.S. Supreme Court declares
2013
genes unpatentable.
Modifying DNA
• Recombinant DNA technology adds genes
from one type of organism to the genome of
another.
o First gene modification biotechnology,
What Is Patentable? and was initially done in bacteria to
• To qualify for patent protection, a transgenic produce peptides and proteins useful as
organism must be new, useful, and non- drugs.
obvious. Recombinant DNA Technology
• Patent law has had to evolve to keep up with • Recombinant DNA technology is also known
modern biotechnology. as gene cloning.
• A DNA sequence alone does not warrant • It began in 1975 when molecular biologists
patent protection. convened to discuss the safety and
o It must be useful as a tool for research or implications of this new technology.
as a novel or improved product, such as a
diagnostic test or drug. • However, it turned out to be safer than
Technology Timeline expected.
Patenting Life and Genes o It also spread to industry faster and in
more diverse ways than imagined.
With gene and genome discoveries
Creating Recombinant DNA Molecules
pouring into the Patent and • Manufacturing recombinant DNA requires
2000 Trademark Office, requirements for restriction enzymes that cut donor and
showing utility of a DNA sequence are recipient DNA at the same sequence.
tightened. • These enzymes cut DNA at sites that are
palindromic.
CYTOGENETICS
• The cutting action of many of these enzymes Recombinant DNA

generate single-stranded extensions called
“sticky ends.”
• Another “tool” used is a cloning vector
• Carries DNA from the cells of one species
into the cells of another
• Commonly used vectors include:
o Plasmids
o Bacteriophages
o Disabled retroviruses
• Cut DNA from donor and plasmid vector with
the same restriction enzyme 1. DNA isolated from donor cell (animal or
• Mix to generate recombinant DNA molecule plant)
• When such a modified plasmid is introduced • A specific restriction enzyme fragments
into a bacterium, it is mass-produced as the donor DNA.
bacterium divides 2. Plasmid isolated from bacterium
Recombining DNA Uses Restriction • The same restriction enzyme that
Enzymes fragmented donor DNA is also used to
Recombining DNA Uses Restriction open plasmid DNA.
Enzymes to Insert a Foreign DNA 3. Donor and plasmid DNA are mixed; “sticky
Sequence ends” of donor DNA hydrogen bond with
sticky ends of plasmid DNA fragment;
recombinant molecule is sealed with ligase.
4. Modified plasmids (recombinant DNA) are
introduced into bacteria.
5. Bacteria divide and clone the gene spliced
into plasmids.
6. Drug is purified and produced.
Selecting Recombinant Molecules
• Three types of recipient cells can result from
attempt to introduce a DNA molecule into a
bacterial cell.
o Cells that lack plasmids
o Cells with plasmids that do not contain
foreign genes
o Cells that contain plasmids with foreign
1. Restriction enzymes cut DNA at specific genes
sequences. Applications of Recombinant DNA
2. DNAs from two sources cut with the same • Recombinant DNA is used to:
restriction enzyme have “sticky” ends. Two o Study the biochemical properties or
pieces of DNA hydrogen bond, and ligase genetic pathways of that protein
seals the sugar- phosphate backbone. o Mass-produce proteins (e.g., insulin)
3. Recombinant DNA molecule. • Sometimes conventional methods are still the
better choice because of economics
CYTOGENETICS
• Textile industry can produce indigo dye in treatment of type

E. coli by genetically modifying genes of the Drug Use
1 diabetes
glucose pathway and introducing genes from Treat genital warts;
another bacterial species hairy cell leukemia;
Interferons hepatitis B and C;
Drugs Produced Using Recombinant DNA Kaposi sarcoma;
Technology multiple sclerosis
Drug Use
Dilates blood
Atrial natriuretic peptide vessels, promotes Treats kidney cancer
Interleukin-2
urination recurrence
Help restore bone Helps lung alveoli
marrow after Lung surfactant to inflate in infants
marrow transplant; protein with respiratory
Colony- stimulating factors restore blood distress syndrome
cells following Renin inhibitor Lowers blood pressure
cancer
chemotherapy Decreases growth in
Somatostatin muscle and bone
Thins secretions
in pituitary gigantism
in lungs of people
Deoxyribonuclease (DNase) Prevents further
with cystic
fibrosis damage to heart
Superoxide dismutase
muscle after heart
Accelerates
attack
healing of
Epidermal growth factor wounds and Stops
Thrombin
burns; treats postsurgical bleeding
gastric ulcers Dissolves blood clots
Stimulates Tissue in treatment of heart
production of red plasminogen activator attack, stroke, and
Erythropoietin (EPO) pulmonary embolism
blood cells in
cancer patients
Promotes blood Transgenic Organisms
clotting in • An even more efficient way to express some
Factor VIII
treatment of recombinant genes is in a body fluid of a
hemophilia A transgenic animal.
Corrects enzyme • Transgenic sheep, cows, and goats have all
Glucocerebrosidase deficiency in expressed human genes in their milk.
Gaucher disease o Clotting factors
Promotes growth o Clot busters
of muscle and o Collagen
bone in people o Antibodies
Human growth hormone with very short • Several techniques are used to insert DNA
stature due to
into cells to create transgenic animals:
hormone
deficiency o Chemicals that open transient holes in
plasma membrane
Allows cells to
Insulin take up o Liposomes that carry DNA into cells
glucose in
CYTOGENETICS
Transgenic Animals o Field tests may not adequately predict the

• Finally, an organism must be regenerated effects on ecosystems
from the altered cell. o Lead to genetic uniformity
• If the trait is dominant, the transgenic animal Some Genetically Modified Organisms
must express it in the appropriate tissue at
the right time in development. Organism Altered Trait
• If the trait is recessive, crosses between Grapes Less sugar
heterozygotes may be necessary to yield Cassava, papaya,
Resist viral infection
homozygotes that express the trait. plum
Animal Models Cattle Resist mad cow disease
• Transgenic animals are far more useful as Sugar beets, corn,
Tolerate an herbicide
models of human diseases. soybeans
o Example: Inserting the mutant human Bananas, rice More iron and vitamin A
beta globin gene that causes sickle cell Altered fatty acid
Canola
anemia into mice composition
• Drug candidates can be tested on these Salmon Faster growth
animal models before testing on humans.
o Will be abandoned if they cause Bioremediation
significant side effects • Transgenic organisms can provide processes
• Transgenic animal models have limitations: as well as products
o Researchers cannot control where a • Bioremediation: The use of bacteria or
transgene inserts, and how many copies plants to detoxify environmental pollutants
do so. • Examples:
o The level of gene expression necessary o Nickel-contaminated soils
for a phenotype may differ in the model o Mercury-tainted soils
and humans. o Trinitrotoluene (TNT) in land mines
o Animal models may not mimic the human Monitoring Gene Function
condition exactly because of differences • Microarrays (gene chips) are devices that
in development or symptoms. detect and display the mRNAs in a cell.
o Piece of glass or plastic that is about 1.5
centimeters square
• Many small pieces of DNA of known
sequence are attached to one surface, in a
grid pattern.
• In many applications, a sample from an
abnormal situation is compared to a normal
control.
Genetically Modified Foods • Messenger RNAs are extracted from the
• Form of genetic modification based on samples and cDNAs are made.
phenotype, such as taste or appearance • These are differentially-labeled and then
• Genetically modified organisms—Organisms applied to the microarray.
altered to have genes from other species or • The pattern and color intensities of the spots
to over- or under express their own genes indicate which genes are expressed.
• Objections:
CYTOGENETICS
• A laser scanner detects and computer 1. Label probes with fluorescent tags.
algorithms interpret the results. 2. Incubate labeled cDNAs with DNA
microarray.
3. Laser scanned detect bound, fluorescent
DNA probes.
4. Computer analyzes data.
Gene Expression Profiling Chronicles
Repair after Spinal Cord Injury
Time After Type of Increased Gene

Injury (rats) Expression
Protective genes to preserve
Day 1
remaining tissue
Day 3 Growth, repair, cell division
1. Isolate RNA Day 10 Repair of connective tissues

2. Generate cDNAs Angiogenesis
Gene Expression in Response to Spinal
Cord Injury Day 30-90 Blood vessel mature
A DNA Microarray Experiment Reveals New type of connective
Gene Expression in Response to Spinal tissue associated
Cord Injury with healing
Gene Silencing and Genome Editing

• Gene silencing techniques block synthesis
of, or degrade, mRNA.
• Genome editing techniques create double-
stranded breaks in the DNA double helix,
enabling insertion of a desired DNA
sequence or removal of a sequence.
Gene Silencing by Antisense Technology

• Block gene expression by introducing RNA
that is complimentary to the gene’s mRNA
transcript.
CYTOGENETICS
• The introduced RNA, called antisense RNA, • Another part of the RISC, a protein
binds to the mRNA, preventing its translation component, acts as a nuclease enzyme
into a protein. degrading targeted RNA, preventing its
Gene Silencing by Use of Morpholinos translation into a protein.
• A morpholino consists of 25 DNA bases RNA Hairpins (Double-Stranded RNA)
bonded to each other by organic groups that • RNA molecules can fold into short, double-
are not the sugar-phosphate ones in DNA. stranded regions where the base sequence
• The morpholinos can block splice-site is complementary.
mutations that would delete exon sites in a
gene.
• One use is in Duchenne muscular dystrophy.
• The morpholino blocks an exon site
necessary to produce effective dystrophin.
Gene Silencing by Use of Ribozyme
• Ribozyme is a RNA-based enzyme in the RNA Interference
ribosome.
• It fits the shapes of certain RNA molecules,
and can cut the RNA molecule.
• Because it can cut the RNA molecule, it
could used to destroy RNA from pathogens,
such as HIV.
Gene Silencing by Use of RNA interference
(RNAi)
• Andrew Fire and Craig Mello won the Nobel
Prize in Physiology or Medicine for explaining
how RNAi works.
• Short double-stranded RNAs sent into cells
separate into single strands, one of which
binds to its compliment in mRNA, preventing
it from being translated.
• RNAi involves the use of double-stranded
RNA and several proteins. 1. Dicer cuts double- stranded RNAs.
• An enzyme termed “dicer” cuts double 2. Double- stranded RNAs separate and bind
stranded RNA in to pieces of 21 to 24 RISC and target mRNA.
nucleotides. 3. Target mRNA cut.
• These short segments of double-stranded Genome Editing
RNA attach to protein complexes, termed • Genome editing uses restriction
RNA-induced silencing complex (RISC). endonucleases to cut and paste DNA
• One strand of the double-stranded short molecules in patterns that might not exist in
RNA, called the guide strand finds attaches nature.
to the protein complex of the RISC. • This technology can be used on somatic cells
• As part of the RISC, the guide strand finds its or germline cells (developing oocytes and
complimentary RNA and binds. sperm).
CYTOGENETICS
• The three major genome editing techniques

are ZFNs, TALENs, and CRISPER-Cas9.
Zinc Finger Nuclease Technology (ZFN)
• This technology uses protein motifs (parts of
proteins that have characteristic shapes)
called zinc fingers.
• A zinc finger is a protein consisting of a beta-
pleated sheet linked to an alpha helix by a
zinc atom. Applications of CRISPR-Cas9 Genome
• Different zinc fingers bind different three- Editing
base DNA sequences.
• If the zinc finger binds, then a nuclease cuts Application Example
the DNA. Deleting genes that encode
hormones that catfish
Transcription-Activator-Like Effector
require to reproduce, so
Nuclease (TALEN) Technology
that catfish with
• In this technology, a restriction enzyme from GMO
other genetic modifications
a bacterium (Xanthomonas) that infects containment
can be raised on
plants cuts DNA on both strands. farms where the hormones
Clustered Regularly Interspaced Short are added to the water, but
Palindromic Repeats (CRISPRs) not survive in the wild.
• These are short sequences of DNA that Replace genes with
include several repeats. Bringing back counterparts from extinct
• They are natural components of the extinct species relatives, such as mammoth
genomes of certain bacteria, where they genes in an elephant.
provide an action similar to an immune Introduce genes into
Limiting spread
disease vectors such as
response. of infectious
mosquitoes and ticks
• CRISPRs enable bacteria to deploy a diseases
that make them sterile.
restriction enzyme called Cas9.
Remove the gene that
• Cas9 enzyme recognizes and cuts out DNA encodes albumin, the egg
sequences from infecting viruses that have Avoiding allergy
white protein, which makes
to vaccines
inserted into the viral genome. some people react to
• The CRISPR DNA sequences along with the vaccines grown in hen eggs.
viral sequences are transcribed into CRISPR Swapping in
RNAs. a CCR5 mutation to provide
Adding genes to
• These RNA serve as guides that bring Cas9 resistance to HIV (see
resist disease
to other sites in the bacterial genome where the chapter opener of
chapter 17).
viral DNA has inserted and then cuts both
strands. Introduce human genes into
pig genomes to create cell
• CRISPR-Cas9 is very versatile; researchers Creating organ
surfaces that the human
can design and synthesize guide RNAs to donors
immune system will not
direct which DNA sequences are removed. reject.
• New DNA sequences can be placed into the Introduce genes for valued
Adding traits to
gene and even new genes can be added. traits (such as fur color,
show animals
body size, and stamina)
CYTOGENETICS
into pets or show animals in

just one or two generations.
Gene Drive
• Gene drive is the application of genome
editing to kill, alter, or render infertile an
pathogen.
• It is based on a natural form of DNA repair,
called homing.
• Homing is a process which removes one
copy of a pair of alleles of a selected gene
and replaces it with another copy of the
remaining allele.
CYTOGENETICS
GENETIC TESTING AND TREATMENT • Genetic counseling sections

Gene Therapy to Genome Editing o Family history
• Gene therapy using genome editing is o Pedigree construction
becoming a valuable medical treatment tool. o Information provided on specific diseases,
• One example of this therapy is for the modes of inheritance, tests to identify at-
treatment of mucopolysaccharidosis type III, risk family members
more commonly known as Sanfilippo o Testing arranged, discussion of results
syndrome type A. and treatments
Genetic Counseling o Links to support groups, appropriate
• Addresses medical, psychological, services, clinical trials
sociological, and ethical issues. o Follow-up contact
Genetic Testing
• Genetic counselor is a health care
professional with a master’s degree who • Genetic tests are administered at all stages
helps patients and their families navigate the of human existence, and for a variety of
confusing path of genetic testing. reasons.
• Genetic counseling began in pediatrics, and • Identifying mutations can help in diagnosis
prenatal care, and has become specialized. and choosing treatments.
• The field has even infiltrated public policy as • As the pace of exome and genome
genetic testing has become widespread. sequencing accelerates, and such testing
Genetic Counselors becomes widely available.
Prenatal
• The U.S. only has about 3000
• Provide information to individuals, couples
Cha
expecting children, and families about: Test Description
pter
o Modes of inheritance
Enriches for X- or Y-
o Disease risks and symptoms
Sperm bearing sperm, then
o Available tests and treatments 6,21
selection intrauterine insemination o
• Interpret direct-to-consumer genetic tests r in vitro fertilization
and assist other health care professionals
Sequences genes, exomes,
with genetic information in their practices and genomes and
• When genetic counseling began, it was Preimplantati examines chromosomes of
“nondirective” on genetic 1 cell of 8-celled embryo
o The practitioner did not offer an opinion or diagnosing a conceived in vitro; 21
suggest a course of action, but presented nd remaining embryo
options sequencing develops for a few days
and is then implanted in
• A more recent definition of the role of the
the uterus
genetic counselor is “shared deliberation and
Chorionic Tests DNA and
decision making between the counselor and
villus samplin chromosomes of chorionic 13
client.” g villus cell
The Genetic Counseling Process
Detects small deletions
• Reasons to seek genetic counseling: and duplications in
o Family history of abnormal chromosomes Rescue
archived cells from past 13
o Elevated risk of single-gene disease karyotyping
unexplained pregnancy
o Family history of multifactorial disease losses
o Family history of cancer
CYTOGENETICS
Noninvasive Adults
Tests cell-free fetal
prenatal 13
(placental) DNA
diagnosis Chapte
Test Description
Maternal Measure levels of r
serum marke biomarkers in pregnant 13 Preconception
rs woman's blood carrier tests for
Tests DNA and Dor Yeshorim progra genetic diseases
Amniocentesi 15
chromosomes of 13 m more prevalent
s
amniocytes among people of
Jewish ancestry
Newborns Tests athletes to
identify carriers
Sickle cell disease 11,12
at risk for
Test Description Chapter symptoms
Screen metabolites Preconception
and DNA in heelstick Comprehensive carri carrier tests for
20
Screening blood sample for 50- 20 er testing many single-
plus actionable gene diseases
conditions Tests for
Sequence genomes of heterozygotes
Genome for diseases
many newborns to 20
sequencing Population
assess clinical value more prevalent 15
carrier screen
in
Children certain populati
on groups
Y chromosome
Cha and
Test Description
pter mitochondria!
Detects small DNA sequences
deletions, identify paternal
Ancestry testing 16
duplications, and and maternal
Chromosomal micro lineages; these
other copy 8
array analysis and autosomal
number variants
associated with markers identify
certain phenotypes distant cousins
Diagnoses Copy numbers
unrecognized of short tandem
syndromes or atypic repeats (STRs)
Forensics testing 14
al cases; family in crime scene
Exome sequencing comparisons 1,4,8 or disaster
distinguish de novo evidence
from inherited
mutations in Test Description Chapter
children Identify remains;
risk for
Military 1
depression,
PTSD; rapid
CYTOGENETICS
infection • Some states perform DNA tests on newborns

diagnosis as well as biochemical tests.
BRCA cancers,
Susceptibility 5,18
Alzheimer disease
Drug efficacy,
Pharmacogenetics adverse effects, 20
and dose
Identify genes
Genome-
contributing small
wide association 7
degrees to a
studies
phenotype
Half of a child's
Paternity genome comes 14 Genetic Testing of Children
from the father
• Chromosomal microarray (CMA) test
Identify gene
detects very small deletions and duplications
variants that
Centenarians 3 that are associated with:
extremely old
people share o Autism
o Developmental delay
o Intellectual disability
Posthumous
o Behavioral problems
o Other phenotypes
Test Description Chapter Genetic Testing of Adults
Identify mutations in • Like children, adults take single-gene tests
Disease
cells of deceased 2,22
diagnosis as part of diagnostic workups based on
individuals
symptoms or other test results.
History Identify remains 5,14
• Faster DNA sequencing has lowered the cost
Compare genomes of
Human of carrier tests to the point that it may be
modern and archaic 16
origins more economical to test everyone for many
humans
diseases with one blood sample per person
Newborn Screening Direct-To-Consumer Genetic Testing (DTC)
• Usually tests for inborn errors of metabolism • Companies market DNA-based tests for
traits, susceptibilities, and genetic diseases
• Are not genetic tests, but instead they use
to the general public.
tandem mass spectrometry to identify
unusual metabolites or chemical imbalances • The Clinical Laboratory Improvement
that indicate a certain disease Amendments (CLIA) control genetic testing.
• The field of newborn screening began in • DTC tests presented as information, not
1961 with the Guthrie test for diagnoses, may not be regulated.
phenylketonuria (PKU). • Unawareness of incomplete penetrance is
o It detects phenylalanine, which builds up one complication of DTC genetic testing.
in affected individuals. • Three Direct-to-consumer tests that provide
• In 1963, a specialized diet became available. information, but not a diagnosis are:
o Tests for inborn athletic ability
• The diet is difficult to follow, but does prevent
o Nutrigenetic testing
mental retardation.
o Pharmacogenetic and pharmacogenomic
• After diet’s success, genetic tests expanded. tests
CYTOGENETICS
Tests for Inborn Athletic Ability activity, decreas

• Several dozen companies worldwide offer ed free radicals
DTC genetic tests for athletic ability. Immune
• The genes tested for are related to fitness response and
IL6 Interleukin-6
and athletic ability. inflammation in
injury repair
• Two genes of particular interest are the
angiotensin-1-converting enzyme gene and Peroxisome
proliferator- Fat storage and
the ACTN3 (alpha-actinin 3) gene.
PPAR activated glucose
Genes Associated With Athletic receptor metabolism
Characteristics gamma
Thyrotropin-
Gene abbrevia Encoded Protein releasing Controls
tion protein Function TRHR
hormone rec metabolic rate
Regulates blood eptor
Beta-2
glucose Degree of
ADRBR adrenergic
level, vasodilatio muscle growth
receptor
n, fat utilization and
Weight gain with Vitamin D
Apolipoprot VDR bone density
APOA2 high saturated receptor
ein A2 increase
fat diet with strength tr
Increased need aining
for Increased blood
Type 1 Vascular
hydration, decre vessel
COL1A1 collagen VEGF endothelial
ased response extensions with
alpha-1 growth factor
to endurance act exercise
ivity
Type 5 Nutrigenetics Testing
Susceptibility to
COL5A1 collagen
tendon injury • Some DTC testing company websites offer
alpha-1
genetic tests along with questionnaires about
Increases fatty
diet, exercise, and lifestyle habits.
acid uptake in
small intestine, • The company then sends a “nutrigenetics”
Fatty acid increasing blood profile with dietary suggestions and pitches
FABP2 binding levels to purchase supplements.
protein 2 of triglycerides, • However, many of these companies provide
LDL, and total inaccurate information.
cholesterol and o Some suggestions are even dangerous.
lowering HDL Matching Patient to Drug
Increases risk of •
Growth A pharmacogenetic test detects a variant of
tendon disease
GDF5 differentiati a single gene that affects drug metabolism.
and osteoarthriti
on factor 5 • A pharmacogenomic test detects variants
s
of multiple genes or gene expression
Gene abbrevi Encoded Protein
patterns that affect drug metabolism.
ation protein Function
Glutathione S Detoxification,
GST
transferase antioxidant
CYTOGENETICS
Pharmacogenetics o Replacing an affected body part or

biochemical with material from a donor
Gene abbr Encoded Geno Phenotyp o Delivering pure, human proteins derived
eviation protein type e from recombinant DNA technology to
Poor compensate for the effects of a mutation
metabolizer o Refolding correctly a misfolded protein
; o Gene therapy, to replace mutant alleles
increased s Treating the Phenotype
Cytochrome
CYP2D6 2D6 ensitivity • Lysosomal storage diseases are a subclass
P450
to certain of inborn errors of metabolism.
beta
• Treatments are based on understanding
blocker hea
rt drugs metabolic pathways.
Strong
o If any enzyme is deficient or its activity
response to blocked, the substrate builds up and the
Serotonin recep G SSRI drugs product is deficient.
HTR2A
tor allele to treat de • There are three general approaches for
pression counteracting these diseases.
and anxiety Counteracting a Metabolic Abnormality
Requires
A118
higher
G
doses of
OPRM1 Opioid receptor (asp
opiates
to asn
for pain reli
)
ef Lysosomal Storage Disease Treatments
4.5-fold
increased
Solute
T/C risk of
carrier organic Treatment Mechanism
SLCO1B1 (val/a muscle pai
anion transport
la) n
er
from statin Recombinant human
drugs enzyme infused
Enzyme replacement
17-fold to compensate for
therapy
increased deficient or
C/C risk of absent enzyme
(ala/a muscle pai Oral drug that reduces
la) n Substrate reduction level of substrate so
from statin therapy enzyme can function
drugs more effectively
Increased
Vitamin K
sensitivity Oral drug that
epoxide reduct 1639
VKORC1 to warfarin Pharmacological bind to patient's
ase complex G>A
(a blood misfolded protein,
subunit 1 chaperone therapy
thinner)
restoring function
Treating Genetic Disease

• Treatments have evolved through stages:
o Removing an affected body part
CYTOGENETICS
Gene Therapy
• Delivers working copies of genes to specific
cell types or body parts, typically aboard
modified viruses
• The first efforts focused on inherited
disorders with a known mechanism, even
though the conditions are rare
• Targeting more common illnesses, such as
heart disease and cancers
Muscle
• Germline gene therapy:
o Gamete or zygote alteration; heritable; • Immature muscle cells (myoblasts) given
not done in humans; creates transgenic healthy dystro-phin genes may treat
organisms muscular dystrophy
• Somatic gene therapy:
o Corrects only the cells that a disease
affects; not heritable
Invasiveness of Gene Therapy
• Ex vivo gene therapy is applied to cells
outside of body that are then returned.
• In vivo gene therapy is applied directly to an
interior body part.
• The most invasive
Gene Therapy Invasiveness
Liver
• To treat certain inborn errors of metabolism,
only 5 percent of the liver’s 10 trillion cells
need to be genetically altered.
Lungs
• Gene therapy can reach damaged lungs
Some Sites of Gene Therapy through an aerosol spray. Enough cells
Endothelium would have to be reached to treat hereditary
• The tile-like endothelium that forms emphysema(alpha-1-antitrypsin deficiency)
capillaries can be genetically altered to or cystic fibrosis
secrete proteins into the circulation
CYTOGENETICS
Somatic Gene Therapy Targets

• Endothelium – Can secrete needed proteins
directly into bloodstream
• Muscle – Accessible, comprises ½ body
mass and has a good blood supply
• Liver – Any functions and can regenerate
• Lungs – Are easily accessed with aerosol
spray
CYTOGENETICS
GENETIC DISORDER • Over 4000 human diseases caused by single

Introduction gene defects
• A genetic disorder is an illness caused by • Examples
one or more abnormalities in the genome, o Adenosine deaminase deficiency, Alpha-1
especially a condition that is present from Antitrypsine deficiency, Cystic fibrosis,
birth (congenital) Galatosemia, Huntington's diseases,
• Most genetic disorders are quite rare and Maple syrup urine disease,
affect one person in every several thousands Phenylketonurimea c, Severe combined
or millions immunodeficiency, Sickle cell disease,
• Genetic disorders may or may not be Smith-Lemil-Optiz Syndrome
heritable, i.e., passed down from the parents' Cystic Fibrosis
genes • Cystic fibrosis is a genetic disorder that affect
Non-heritable the respiratory and digestive systems
• In non-heritable genetic disorders defects • People with cystic fibrosis inherit a defective
may be caused by new mutations or changes gene on chromosome 7 called CFTR (cystic
to the DNA fibrosis transmembrane conductance
• The defect will only be heritable if genetic regulator)
disorder occurs in the germ line • The protein produced by this gene normally
Facts helps salt (sodium chloride) move in and out
Earliest Evidence of Hereditary Genetic of cells
Disorder Discovered • If the protein doesn't work correctly, that
movement is blocked and an abnormally
thick sticky mucus is produced on the outside
of the cell
• The cells most seriously affected by this are
the lung cells. This mucus clogs the airways
in the lungs, and increases the risk of
infection by bacteria
• The thick mucus also blocks ducts in the
pancreas, so digestive enzymes can't get into
the intestines
• The fossil is dated as 1.5 million years old
and is from a two-year old Homo erectus
child. Homo erectus was a precursor of
modern man
Classification
• Single gene disorder
• Chromosomal genetic disorder
• Multifactorial genetic disorder
Single Gene Disorder
• These disorders involve mutation in the DNA Galactosemia
sequences of single genes. As a result, the • Galactosemia is a rare disorder that affects
protein the gene codes for is either altered or the body's ability to break down a food sugar
missing called galactose
CYTOGENETICS
• The body breaks down lactose into galactose • Some of infections including pneumonia
and glucose and uses these sugars for (lung infection). meningitis (brain infection),
energy and sepsis (blood infection)
• Most people with galactosemia are missing Sickle Cell Disease
an enzyme (called GALT) that helps further • Sickle cell disease is a disorder that affects
break down galactose the red blood cells, which use a protein
• Defects in galactose metabolism cause toxic called hemoglobin to transport oxygen from
chemicals to build up in cells of the body the lungs to the rest of the body
• Normally, red blood cell are round and
flexible so they can travel freely through the
narrow blood vessels
• The hemoglobin molecule has two parts: An
alpha and a beta
• Patients with sickle cell disease have a
mutation in a gene on chromosome 11 that
codes for the beta subunit of the hemoglobin
protein
Sever Combined Immunodeficiency (SCID) • As a result, hemoglobin molecules don't form
• SCID is a group of very rare and potentially properly, causing red blood cells to be rigid
fatal-inherited disorders related to the and have a concave shape (like a sickle)
immune system
• People with SCID have a defect in their
immune system that leaves them vulnerable
to potentially deadly infections
• The most common form is caused by a
mutation in the SCIDX1 gene located on the
X chromosome
• This gene encodes a protein that is used to
construct a receptor called IL2RG (interleuin-
2 receptor)
• These receptors reside in the plasma
membrane of immune cells
• Their job is to allow two types of immune
cells - T cells and B cells to communicate
• When the gene is mutated, the receptors
cannot form and are absent from immune
cells Fibrodysplasia Ossificans Progressiva
• As a result, the immune cells can't (FOP)
communicate with one another about • Sometimes called "stone man" syndrome, it
invaders in the environment. Not enough T is extremely rare genetic disease causes soft
and B cells are produced to fight off the tissue to turn into bone
infection, and the body is left defenseless
CYTOGENETICS
length of which may vary) on the short arm of

chromosome 5
• Multiple genes are missing as a resulr of this
deletion, and each may contribute to the
symptoms of the disorder
• Babies with cri-du-chat are usually small at
birth, and may have respiratory problems
• Often, the larynx doesn't develop correctly,
which causes the signature cat-like cry
Chomorsomal Genetic Disorder

• In these disorders, entire chromosomes, or
large segments of them, are missing,
duplicated, or otherwise altered
• Can be organized into two basic groups:
1. Numerical Abnormalities: When an individual
is missing either a chromosome from a pair
(monosomy) or has more than two
chromosomes of pair (trisomy)
2. Structural Abnormalities: When the
chromosome's structure is altered Down Syndrome
• Examples: • Down syndrome is a developmental disorder
o Cri-Du-Chat Syndrome. Down Syndrome. caused by an extra copy og chromosome 21
47 XXY Klinefelter Syndrome, Turner (which is why the disorder is also called
Syndrome (Monosomy), William "trisomy 21")
Syndrome • Having an extra copy of this chromosome
means that individual have three copies of
each of its genes instead of two, making it
difficult for cells to properly control how much
protein is made
• Producing too much or too little protein can
have serious consequences
• People with down syndrome have very
distinct facial features: A flat face, a small
broad nose, abnormally shaped ears, a large
Cri-Du-Chat Syndrome
tongue, and upward slanting eyes with small
• The name of this syndrome is French for "cry folds of skin in the corners
of the cat," referring to the distinctive cry of
• People with down syndrome have an
children with this disorder
increased risk of developing a number of
• The cry is caused by abnormal larynx medical problems: Respiratory infections,
development, one of the many symptoms gastrointestinal tract obstruction, leukemia,
associated with this disorder heart defects, hearing loss, hypothyroidism,
Cri-du-char is caused by a deletion (the and varies eye abnormalities
CYTOGENETICS
• No cure exists for down syndrome Turner Syndrome

• But physical therapy and/or speech therapy • Turner syndrome is cause by a missing or
can help people with the disorder develop incomplete X chromosome
more normally • People who have turner syndrome develop
as females
• The genes affected are involved in growth
and sexual development, which is why girls
with the disorder are shorted than normal
and have abnormal sexual characteristics
William Syndrome
• Williams syndrome is a rare genetic disorder
that affects a child’s growth, physical
appearance and cognitive development
• People who have William's syndrome are
missing genetic material from chromosome
7, including the gene elastin
47, XXY (Klinefelter Syndrome)
• Males (46, XY) and females (46, XX)
• 47, XXY or ( XXY) is a genetic condition
caused when someone has two X
chromosomes and one Y chromosome
• Because people with an XXY chromosome
arrangement have a Y chromosome, they are
considered genetic males
• Most XXY individuals develop as males,
often not knowing they have an extra
chromosome
• XXY is usually caused by what is called Colorblindness
nondisjunction • Mutated genes are located on the X-
• Nondisjunction happens when a pair of sec chromosome (for red/green colorblindness or
chromosome fails to separate during egg (or both the X and Y chromosome (for total
sperm) formation colorblindness)
• The XXY chromosome arrangement affects
primarily sexual development)
• are lower than average
• As adults, nearly all XXY males are unable to
make sperm and so cannot have biological
children
• Many men discover their condition only after
they seek medical help for infertility
• Changes that appear at puberty can include
low growth of facial and body hair,
development of breast tissue, and small
testes
CYTOGENETICS
Multifactorial Disorder • These plaques and tangles damage the

• These disorders involve variations in multiple healthy cells around them
genes, often coupled with environmental • Because Alzheimer's destroys brain cells,
causes people who have the disorder slowly lose
• Example their ability to think clearly
o Alzheimer's Disease • At first, they may forget words or names, or
o Breast/Ovarian Cancer have trouble finding things
o Colon Cancer • As the disorder worsens, they may forget
o Hypothyroidism how to do simple tasks
o Asthma
o Cancers
o Cleft Palate
o Diabetes
o Heart Disease
o Hypertension
o Inflammatory Bowel Disease
o Mood Disorder
o Obesity
o Refractive Error
o Infertility
Alzheimer's Disease
• Alzheimer's is a disease that causes
dementia or loss of brain function. It affects
the parts od the brain that deal with memory,
thought, and language
• The brain of a person with Alzheimer's
contains abnormal clumps of cellular debris
and protein (plaques) and collapsed
microtubules (support structure of the cell)
• Microtubule disintegration is caused by a

malfunctioning protein called tau, which
normally stabilizes the microtubules
• In Alzheimer's patients, tau proteins instead
cluster together to form disabling tangle

Cytogenetics

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BACHELOR OF SCIENCE IN MEDICAL TECHNOLOGY

NATIONAL UNIVERSITY – MALL OF ASIA

THE GENETIC MATERIAL o Thus, DNA is the genetic material

James Watson and Francis Crick o A phosphate group

Investigator Contribution Timeline

• Nontemplate strand of the DNA double helix

3. Deoxyribose as the 3. Ribose as the sugar

o Stores RNA- and protein-encoding Associates

Enzymes in DNA Replication

Steps of DNA Replication

• DNA polymerase binds nucleotides to form

8. Enzymes remove RNA primers. Ligase seals

4. DNA polymerase adds DNA nucleotides to

6. Continuous strand synthesis continues in a 5’

1. Select target sequence in virus genome.

Transcription Setting the Stage for Transcription to

Simultaneous Transcription of mRNAs

o Different codons that specify the same

Reading Frame • GGA bonds to its complementary anticodon,

• The ribosomal subunits separate and are o Secondary (2°) structure

1. Primary structure – The sequence of amino

o Stabilize partially folded regions in their Diseases Associated with Protein

Lewy body dementia Alpha synuclein

Prion diseases Prion protein

GENE MUTATION o A person with a somatic mutation has

Oncogene Multiple • Caused by mutagens, many are also

Spontaneous Mutation Rate

• Transversion = Purine replaces pyrimidine Single-base

Not All Mutations Impact Protein Function

DNA TECHNOLOGIES Attempts to enforce patents on non-

• The cutting action of many of these enzymes Recombinant DNA

• Textile industry can produce indigo dye in treatment of type

Transgenic Animals o Field tests may not adequately predict the

Time After Type of Increased Gene

1. Isolate RNA Day 10 Repair of connective tissues

Gene Silencing and Genome Editing

Gene Silencing by Antisense Technology

• The three major genome editing techniques

into pets or show animals in

GENETIC TESTING AND TREATMENT • Genetic counseling sections

infection • Some states perform DNA tests on newborns

Tests for Inborn Athletic Ability activity, decreas

Pharmacogenetics o Replacing an affected body part or

Treating Genetic Disease

Somatic Gene Therapy Targets

GENETIC DISORDER • Over 4000 human diseases caused by single

length of which may vary) on the short arm of

Chomorsomal Genetic Disorder

• No cure exists for down syndrome Turner Syndrome

Multifactorial Disorder • These plaques and tangles damage the

• Microtubule disintegration is caused by a

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