CRISPR                   
    (acronym for clustered
regularly interspaced short
palindromic repeats)

Pentru cine nu stie engleza:Repetări palindromice scurte interspațiate în mod regulat în grup.

         Gene editing
*What is gene editing?
Gene editing is a type of genetic engineering in
which DNA is inserted, deleted, modified or replaced in
the genome of a living organism.

*Gene Editing Techniques: Tools to Change The Genome

• Restriction Enzymes: the Original Genome Editor.
• ZFNs (zinc finger nucleases): Increased Recognition
Potential.
• TALENs (transcription activator-like effector
nucleases)Gene Editing: Single Nucleotide Resolution.
• CRISPR-Cas9 Gene Editing: Genome Editing
Revolutionized.
*Why CRISPER-CAS9 revolutionized genome editing?
For this we need to look at a brief history of genome
engineering.

Smart people ask questions!!!

Genome Editing: Past, Present, and Future
Past: Classically, genetic studies relied on the discovery
and analysis of spontaneous mutations .In the mid-
twentieth century, Muller and Auerbach demonstrated
that the rate of mutagenesis could be enhanced with
radiation or chemical treatment .The first targeted
genomic changes were produced in yeast and in mice in
the 1970s and 1980s.This gene targeting depended on
the process of homologous recombination, which was
remarkably precise but very inefficient, particularly in
mouse cells. Recovery of the desired products required
powerful selection and thorough characterization.

Present: The secret to high-efficiency genome editing is the

ability to make a targeted DNA double-strand break (DSB) in
the chromosomal sequence of interest .We are currently
endowed with three powerful classes of nucleases that can
be programmed to make DSBs at essentially any desired
target and we will talk about them in the next
presentation.
Future of genetic engineering
• Germline editing. Stimulated by recognition of the ease of CRISPR-based editing and the possibility of
misuse of the technology, there is considerable current interest in prospects for human germline
genome editing. Such applications would involve delivery of the editing reagents to embryos created
by in vitro fertilization. In the future, it may be feasible to engineer gametogenic precursor cells in
prospective parents instead. The advantage to germline correction of disease alleles is that they will
forever be gone from the lineage of the treated individual. The risk at present is that the attempt to
correct may do more harm than good. Current genome editing technology does not have sufficient
efficiency and specificity to be reliably safe. Mutations generated at non-target sites in the genome will
also affect the treated person and be transmitted through subsequent generations, and their effects will
not always be benign or predictable, nor will they be readily reversible .In the medical realm, what
therapies will be developed based on whom we decide needs to be “fixed”? Devastating diseases, like
Huntington’s disease and muscular dystrophy, are obvious candidates. What about hereditary deafness
or short stature? People with these conditions are often high-functioning, have strong communities, and
do not feel themselves to be in need of “correction” . To take an absurd example, is skin color a
condition that needs altering? This brings us to purely cosmetic changes that some may find desirable –
hair color, eye color, height, athletic ability (assuming we know how to engineer these traits genetically).
Should these applications be pursued?1
• Once methods are developed, who will benefit? Human therapies based on genome editing are
currently complex and expensive. Will only the wealthy be able to afford them?
Advantages of using
CRISPR
• CRISPR not only works with remarkable ease and speed,
but is also the first technique that allows researchers to
modify multiple genes in a single experiment.
• CRISPR-Cas9 is easy to use, especially compared to older
genomic editing tools. "Before, only a small number of
laboratories in the world could produce the right tools for
gene editing. Now, even a high school student can make a
change in a complex genome using CRISPR, ”said Prof. Dr.
Alejandro Chavez of Columbia University, who has
developed several new tools based on CRISPR technology.
• CRISPR is also easy to adapt and customize according to
your goal. It can edit virtually any segment of DNA in the
3 billion letters of the human genome and is more
accurate than other DNA editing tools.
• Genomic editing with CRISPR-Cas9 is much faster. In the
case of older methods, it usually takes at least a year or
two to generate a genetically modified mouse model. But
now, with the help of CRISPR technology, a scientist can
create a complex mouse model in a matter of months.
And in addition, it is certainly cheaper than previous
methods.
How CRISPR works?
Medical applications of CRISPR

CRISPR can cure:

•B-cell Acute Lymphoblastic Leukemia
•Lung and Esophageal cancers
•Multiple Myeloma
•Cervical cancer
•Beta Thalassemia
•Alzheimer’s Disease
•Huntington’s Disease
•Duchenne Muscular Dystrophy
•Retinitis Pigmentosa
•HIV
•Diabetes
•Autism Spectrum Disorder

And so on,for now

they are only in
theory and studies
Topic of
debate

• Taking into account

ethical
considerations.
• What do you think
about designer babies?
Va multumesc!