Professional Documents
Culture Documents
Basic Principles of
Application of Genetic Engineering
as a New Modality of Treatment of
Many Congenital Disorders and
Cancer
Prof. Dr. Mohamed Ali, M.Sc.. M.Phil., B.Ed., M.S (Sing)., Ph.D (Sing)
Department of Clinical Biochemistry
Faculty of Medicine
University of Tabuk
Dr.Padmesh. V
GENETIC ENGINEERING
Changing the genetic information in a cell.
m o d e l o r g a n i s m s
sizes
Dr.Padmesh. V
v i r u s e s
b a c t e r i a
yeast
Saccharomyces cerevisiae
control of cell cycle and cell
division
protein secretion and
membrane biogenesis
function of the cytoskeleton
cell differentiation
aging
gene regulation and
chromosome structure
Dr.Padmesh. V
r o u n d w o r m
Caenorhabditis elegans
development of the body
plane
cell lineage
formation and function of the
nervous system
control of programmed cell
death
cell proliferation and cancer
genes
aging
behaviour
gene regulation and
chromosome structure
Dr.Padmesh. V
f r u i t f l y
Drosophila melanogaster
development of the body plan
generation of differentiated cell
lineages
formation of the nervous system,
heart and musculature
programmed cell death
genetic control of behaviour
cancer genes and control of
cell proliferation
control of cell polarisation
effect of drugs, alcohol and
pesticides
Dr.Padmesh. V
f r u i t f l y
Dr.Padmesh. V
zebrafish
development of
vertebrate body
tissue
formation and
function of brain and
nervous system
birth defect
cancer
Dr.Padmesh. V
mice
development of body
tissues
function of mammalian
immune system
formation and function of
brain and nervous system
models of cancer and
other human diseases
gene regulation and
inheritance
infectious disease
Dr.Padmesh. V
APPLICATIONS OF
GENETIC ENGINEERING
Dr.Padmesh. V
Since their introduction in 1996, genetically modified (GM) plants have become
an important component of our food supply.
One genetic modification uses bacterial genes that produce a protein known
as Bt toxin.
This toxin is harmless to humans and most other animals, but enzymes in the
digestive systems of insects convert Bt to a form that kills the insects.
GM ANIMALS
Transgenic animals are becoming more important to our food
supply.
About 30 percent of the milk in U.S. markets comes from cows that
have been injected with hormones made by recombinant-DNA
techniques to increase milk production.
Milk from these goats may help prevent infections in young children
who drink it.
Dr.Padmesh. V
PREVENTING DISEASE
Two genes engineered into the rice genome help the grains produce
and accumulate beta-carotene.
TREATING DISEASE
Recombinant-DNA technology can be used to make
important proteins that could prolong and even save human
lives.
TREATING DISEASE
TREATING DISEASE —
ONE EXAMPLE OF GENE THERAPY
To deliver therapeutic genes to target cells researchers engineer a virus that cannot
reproduce or cause harm.
The DNA containing the therapeutic gene is inserted into the modified virus.
The patient’s cells are then infected with the genetically engineered virus.
In theory the virus will insert the healthy gene into the target cell and correct the
defect.
Dr.Padmesh. V
TREATING DISEASE
• Gene therapy can be risky.
FORENSIC SCIENCE
PERSONAL IDENTIFICATION
In DNA fingerprinting, restriction enzymes first cut a small sample of human DNA into
fragments containing genes and repeats.
Note that the repeat fragments from these two samples are of different lengths.
DNA samples can be obtained from blood, sperm, or tissue—even from a hair strand
if it has tissue at the root.
Dr.Padmesh. V
Gene therapy is the the delivery of therapeutic gene into a patient's cells
to treat disease.
Cell therapy is the delivery of intact, living cells into a patient to treat
disease.
Alright,tell me
how you want to
change…!
Gene
Dr.Padmesh. V
METHOD:
Introduction of FUNCTIONAL GENES into appropriate cells
Transferred gene (TRANSGENE) encodes & produces proteins
The Proteins encoded by Transgene corrects the disorder
Dr.Padmesh. V
Gene Therapy:
Approaches:
Two ways to deliver genes:
1. Ex vivo approach
2. In vivo approach
Dr.Padmesh. V
1. Ex vivo approach:
-Target cells are removed from the body and grown in vitro.
-The gene is then introduced into the cultured cells.
-These cells are then re-introduced into the same individual
-Examples: Fibroblast cells, Hematopoietic cells.
Dr.Padmesh. V
Dr.Padmesh. V
2. CHEMICAL METHODS:
-Calcium phosphate
-DEAE-Dextran
-Liposomes
3. BIOLOGICAL METHODS:
Viral Vectors like
-Retrovirus -Adenovirus -HSV
Dr.Padmesh. V
4. NEO-ORGAN IMPLANTS
5. TISSUE TRANSPLANTATION
7. OTHERS:
-Receptor mediated delivery
-Virally directed enzyme prodrug therapy
Dr.Padmesh. V
2. GERMLINE THERAPY:
Examples:
1. Deficiency of ADA
2. Haemophilia
Dr.Padmesh. V
Examples:
This strategy is useful in Cancers caused by inappropriate
expression of a gene.
Dr.Padmesh. V
Examples:
1. Introduction of a gene that makes cancer cells
susceptible to anticancer drugs.
2. Introduction of a toxic gene whose expression kills
cancer cells.
3. Genes of cytokines can be introduced into cells of
immune system to enhance their potential to kill
diseased cells.
Dr.Padmesh. V
COMMON VECTORS
USED FOR GENE
THERAPY:
1.Retro viruses
2. Adeno viruses
3. Liposomes
Dr.Padmesh. V
1. RETRO VIRUSES:
Retroviruses used in gene therapy are made incapable of
independent replication,to prevent side effects associated
with infectivity.
Retroviruses are used ONLY in EX VIVO THERAPY.
Advantages:
Chromosomal integration & stable modification of target
cells.
Disadvantages:
Uncontrolled integration; May be oncogenic.
Cannot infect non-dividing cells.
Dr.Padmesh. V
2. ADENO VIRUSES:
Second most commonly used delivery system in gene
therapy.
Adenoviruses can be produced at high titres in cultures.
Advantages:
Can infect non-dividing cells,thus suitable for gene therapy
of Cystic fibrosis, DMD.
Non-integration to chromosome. Avoids the risks of
uncontrolled integration.
Efficient gene transfer.
Disadvantages:
Transient expression of gene due to episomal integration.
Provokes immune response.
Dr.Padmesh. V
Dr.Padmesh. V
3.LIPOSOMES:
These are lipid bilayers surrounding an aqueous vesicle.
Can be used to introduce foreign DNA into a target cell.
Advantages:
Safer when compared to Viral vectors.
Can carry large DNA molecules.
Disadvantages:
Inefficient transfer.
Transient expression.
Dr.Padmesh. V
Dr.Padmesh. V
BLINDNESS
PARKINSON’S DISEASE
Dr.Padmesh. V
HOW IT WORKS??
used harmless viruses
enable access to the cells beneath the
retinas of patients
By using a very fine needle
-safe in an extremely fragile tissue and can
improve vision in a condition previously
considered wholly untreatable
Dr.Padmesh. V
HOW IT WORKS??
Done with local anesthesia, used a harmless,
inactive virus [AAV-2 GAD].
Their HSCs are gene modified to resist HIV, and are then
transplanted back into the participant in a mix of modified
and unmodified cells.
Dr.Padmesh. V