Mutasi dan Rekayasa

Genetika

Gita Vita Soraya, MD PhD

 Mutasi
Sasaran Pembelajaran
1 Menjelaskan jenis-jenis variasi genetik

2 Menjelaskan jenis-jenis mutasi

3 Menjelaskan penyebab mutasi

4 Menjelaskan mekanisme repair

 Jenis variasi DNA

POLIMORFISME
1. Bila terjadi pada >1% populasi maka disebut
polimorfisme
2. Bisa satu nukleotida (SNP) atau beberapa sekuens
repetitive (length polymorphism)
3. Biasanya tidak menyebabkan penyakit secara
langsung, namun bisa jadi faktor predisposisi

MUTASI
1. Bila terjadi permanen sekuens DNA suatu gen dimana
terjadi perubahan pada <1% populasi
2. Secara umum dibagi menjadi:
• Germline (pada seluruh sel, diwariskan)
• Somatic (pada sel tertentu saja, tidak
diwariskan)
 lead to amino acid change and result in
Missense
Jenis-jenis mutasi mutation
production of abnormal protein

do not lead to a change in amino acids

Point mutation Silent mutation

Small-scale when substitution of a single nucleotide

Nonsense
(sequence Deletions results in formation of a stop codon and a
mutation
mutations) truncated protein

Insertions

Mutations Numerical loss or duplication of the entire chromosome

chromosomal
change

translocations or inversions that result in an exchange of chromosomal

Chromosomal segments between two nonhomologous chromosomes
rearrangement
Large-scale
(chromosomal
alterations) when a particular chromosomal region is repeated multiple times on
Amplification the same chromosome or different chromosomes, resulting in the
increased copy number of the gene located within this region

Chromosomal loss of heterozygosity (LOH), when deletion of a discrete chromosomal

deletion (LOH) region leads to loss of a tumor suppressor gene residing in this area
Penyebab Mutasi Causes of
Mutation

Induced Spontaneous
(environmental causes or mutagens) (arise naturally during replication)

Ionizing radiation

Nonionizing
radiation (UV)

Mutagenic chemicals
Penyebab Mutasi Causes of
Mutation

Induced Spontaneous
(environmental causes or mutagens) (arise naturally during replication)

Mechanism:
Ionizing radiation X-rays and nuclear radiation eject electrons from
atoms  electrically charged ions  promote
chemical reactions that change DNA bases or break
the bonds of double-stranded DNA  can reach all
cells of the body, including the germline cells.

Nonionizing Prominent example:

radiation (UV) 1. Fukushima nuclear meltdown: sharp increase in
mortality and non-communicable diseases
2. Chernobyl meltdown: 40x increase of thyroid
cancer in children

Mutagenic chemicals
Penyebab Mutasi Causes of
Mutation

Induced Spontaneous
(environmental causes or mutagens) (arise naturally during replication)

Mechanism:
Ionizing radiation Nonionizing radiation (UV)  move electrons 
forms covalent bonds between pyrimidine bases
(cytosine/thymine)  pyrimidine dimers formed 
failure to pair with purines during replication

Prominent example:
Nonionizing Skin cancer
radiation (UV)

Mutagenic chemicals
Penyebab Mutasi Causes of
Mutation

Induced Spontaneous
(environmental causes or mutagens) (arise naturally during replication)

Ionizing radiation

Nonionizing
radiation (UV)

Mutagenic chemicals
 DNA repair helps to ensure the accuracy of the DNA
sequence by correcting replication errors (mismatches),
Mekanisme Repair repairing double-stranded DNA breaks, and excising
damaged nucleotides.

Base Excision Repair Nucleotide Excision Repair

 Rekayasa Gen
Sasaran Pembelajaran

1 Menjelaskan contoh rekayasa genetika dalam

bidang medis

2 Menjelaskan penggunaan terapi gen untuk mutasi

dan contoh applikasinya
 Rekayasa Genetik dalam Bidang Medis

Rekayasa genetik adalah proses manipulasi atau modifikasi

gen organisme dengan menggunakan bioteknologi seperti:
• DNA rekombinan
• Sintesis DNA artifisial

Penggunaannya dalam kedokteran:

• Terapi gen
• Menghasilkan model hewan (knock out models) yang
digunakan untuk penelitian
• Produksi hormon dan vaksin
Rekayasa Genetik dalam Bidang Medis

Terapi Gen

Terapi gen adalah teknik eksperimental yang menggunakan gen

sebagai terapi atau pencegahan penyakit

Pendekatan terapi gen:

• Mengganti gen yang mutasi (menyebabkan penyakit)
dengan copy yang sehat
• Knock out gen mutasi
• Memperkenalkan gen baru

Untuk memasukkan gen dalam tubuh, dibutuhkan vector yang

biasanya merupakan virus termodifikasi. Retrovirus
menyebabkan integrase gen ke kromosom, dan adenovirus
tidak menyebabkan integrase DNA ke dalam kromosom
Rekayasa Genetik dalam Bidang Medis

Terapi Gen
 Rekayasa Genetik dalam Bidang Medis

Terapi Gen

Several inherited immune deficiencies have been treated

successfully with gene therapy. Most commonly, blood stem
cells are removed from patients, and retroviruses are used to
deliver working copies of the defective genes. After the genes
have been delivered, the stem cells are returned to the patient.
Because the cells are treated outside the patient's body, the
virus will infect and transfer the gene to only the desired target
cells.
Severe Combined Immune Deficiency (SCID) was one of the
first genetic disorders to be treated successfully with gene
therapy, proving that the approach could work. However, the
first clinical trials ended when the viral vector triggered
leukemia (a type of blood cancer) in some patients. Since then,
researchers have begun trials with new, safer viral vectors that
are much less likely to cause cancer.
 Rekayasa Genetik dalam Bidang Medis

Terapi Gen

Patients with beta-Thalassemia have a defect in the beta-globin

gene, which codes for an oxygen-carrying protein in red blood
cells. Because of the defective gene, patients don't have
enough red blood cells to carry oxygen to all the body's tissues.
Many who have this disorder depend on blood transfusions for
survival.
In 2007, a patient received gene therapy for severe beta-
Thalassemia. Blood stem cells were taken from his bone
marrow and treated with a retrovirus to transfer a working
copy of the beta-globin gene. The modified stem cells were
returned to his body, where they gave rise to healthy red blood
cells. Seven years after the procedure, he was still doing well
without blood transfusions.
A similar approach could be used to treat patients with sickle
cell disease
 Rekayasa Genetik dalam Bidang Medis

Terapi Gen

Several promising gene-therapy treatments are under

development for cancer. One, a modified version of the herpes
simplex 1 virus (which normally causes cold sores) has been
shown to be effective against melanoma (a skin cancer) that has
spread throughout the body. The treatment, called T-VEC, uses a
virus that has been modified so that it will (1) not cause cold
sores; (2) kill only cancer cells, not healthy ones; and (3) make
signals that attract the patient's own immune cells, helping them
learn to recognize and fight cancer cells throughout the body. The
virus is injected directly into the patient's tumors. It replicates
(makes more of itself) inside the cancer cells until they burst,
releasing more viruses that can infect additional cancer cells.
A completely different approach was used in a trial to treat 59
patients with leukemia, a type of blood cancer. The patients' own
immune cells were removed and treated with a virus that
genetically altered them to recognize a protein that sits on the
surface of the cancer cells. After the immune cells were returned
to the patients, 26 experienced complete remission.