GENE BASED PRODUCT

ST. LIDETA HEALTH SCIENCE AND BUSINESS COLLEGE

IMMUNOLOGY GROUP ASSIGNMENT
DEPARTMENT OF PHARMACY
SECTION : H
Sub to: Mr. Alemsegd
Sub date: Feb.9, 2024
 GENE BASED PRODUCTS

• Also known as genetic product.

• Gene based product is a fascinating of study that combines
genetics and product development.
• Are goods or services that are derived from genetic
material, genetic manipulation or genetic engineering
process.
• A gene product is the biochemical material,
either RNA or protein, resulting from expression of a gene.
• A measurement of the amount of gene product is
sometimes used to infer how active a gene is.
• A gene is defined as "a hereditary unit of DNA that is
required to produce a functional product"
• examples of gene based products include:
– human recombinant insulin, growth hormone,
blood clotting factors, hepatitis B vaccine, and
diagnosis of HIV infection.
 WHAT IS GENE THERAPY?

• Gene therapy is the insertion , alteration, or removal of genes

within an individuals cells to treat diseases.
• It is a technique for correcting defective genes that are responsible
for disease development.
• There are four approaches
i. A normal gene inserted to compensate for a non functional
gene
ii. An abnormal gene traded for a normal gene
iii. An abnormal gene repaired through selective reverse mutation.
iv. Change the regulation of gene pairs.
HISTORY

• In the early 1970’s scientist proposed gene surgery.

• In 1983, a group of scientist from Baylor college of Medicine

in Houston,Texas, proposed gene therapy for Lesh-Nyhan
disease, a rare neurological disorder.

• On september 14, 1990 a 4 year old suffering from a genetic

disorder that prevented her body from producing a crucial
enzyme became the first person to undergo gene therapy in the
U.S.
TYPES OF GENE THERAPY

GERM LINE GENE THERAPY SOMATIC CELL GENE THERAPY

• Therapeutic genes transferred • Therapeutic genes transferred

into the germ cells. into the somatic cell.
• Eg. Genes introduced into • Eg. Introduction of genes into
eggs and sperm the blood cell , skin cell etc.
• It is heritable and passed on to • Will not be inherited by later
later generation generation
• For safety and ethical reasons • At present all research s
it is not being attempted at directed to correct genetic
present. defects in somatic cell.
 GERM LINE GENE THERAPY

• In the case of germ line gene

therapy, germ cell ,i.e., sperm
and egg are modified by the
introduction of functional
genes.
• Therefore the change due to
therapy would be heritable and
would be passed on to later
generation.
• For ethical reasons it is not
being attempted at present
SOMATIC CELL GENE
THERAPY
EX VIVO GENE THERAPY

• ADVANTAGES

• Possibility to expand
different cell population
• Select cells in which
gene transferred has
occurred
• Avoid possibility of
immune response.
IN VIVO GENE THERAPY
Gene delivery system

• Gene delivery is the process of introducing foreign

genetic material, such as DNA or RNA, into
host cells.
• Gene delivery must reach the genome of the host cell
to induce gene expression.
• Successful gene delivery requires the foreign gene
delivery to remain stable within the host cell and can
either integrate into the genome or replicate
independently of it.
Cont.…

• This requires foreign DNA to be synthesized as part

of a vector, which is designed to enter the desired
host cell and deliver the transgene to that cell's
genome
• Based on Vectors utilized as the method for gene
delivery can be divided into two categories,
– recombinant viruses(viral)
– synthetic vectors (non-viral).
Methods

• There are a variety of methods available to deliver

genes to host cells.
• When genes are delivered to bacteria or plants the
process is called transformation and when it is used
to deliver genes to animals it is called transfection.
• This is because transformation has a different
meaning in relation to animals, indicating progression
to a cancerous state
Cont.…

• For some bacteria no external methods are need to

introduce genes as they are naturally able to take up
foreign DNA.
• Most cells require some sort of intervention to make
the cell membrane permeable to DNA and allow the
DNA to be stably inserted into the hosts genome.
1. Viral gene delivery systems

 Viral gene delivery can be by modifications in

adenoviruses, retroviruses, adeno-associated viruses
and etc. these vectors can be can be classified in to
two:
• Nonlytic: those that produce virions but leave host cell
intact. For example : adenoviruses, lentiviruses
• Lytic: those that produce virions and destroy host cell. For
example human adenoviruses and herpes simplex viruses.
 Viral vector have high transduction efficiency but may pose
safety concerns related to immunogenicity and potential
integration in to the host genome.
RETROVIRAL VECTOR
• The genetic molecule of
retrovirus is RNA.
• Retroviral vectors are created
by removal of the retroviral
gag, pol , env genes. These are
replaced by therapeutic gene.
• When retrovirus infect a host
cell , it produce a DNA copy
from its RNA with the help of
reverse transcriptase.
• DNA is incorporated into the
host genome with the help of
integrase.
• The integrase enzyme can
insert the genetic material in
any arbitrary position.
 ADENO VIRAL VECTOR

• These are non-enveloped , composed

of nucleocapsid and dsDNA genome.
• Viral DNA is removed and
therapeutic gene is inserted.
• They introduced their DNA molecule
into the host.
• The genetic material is not
incorporated into the host cell’s
genetic material.
• The DNA molecule is left free in the
nucleus and the instruction the DNA is
transcribed.
• The extra gene are not replicated when
the cell is about to undergo cell
division.
Adeno-associated viruses

• Small single stranded DNA virus causing mild immune

response in human and other primates derived from
parvoviradae family.
• AAV vectors are used for gene therapy as these affect
only the dividing cells. AAV gene therapy in retina is
almost a success.
• It is 20nm long and 4.5kb size non enveloped DNA
viruses.
• Limited transgene capacity of the particles.
Herpes simplex virus
• HSV system include the development of the so called
disabled infectious single copy viruses, which
comprises a glycoprotein H defective mutant HSV
genome.
2. Non-viral gene delivery system

• Non viral vector offer the advantage of reduced

immunogenicity and potential for large scale
production.
• Simple and safer alternative for the viral system and
low host immunogenicity will be two.
• These vector often rely on cationic polymer or lipids to
condense and protect genetic material for delivery.
• Non viral gene delivery is divided in to two:
 physical(electroporation, gene gun…)
 chemical(liposomes, polymers and etc)
 LIPOSOMAL VECTOR
• A minute spherical sac of
phospholipid molecules
enclosing a water droplet,
artificially formed to carry
drugs or other substances
into the tissues.
• In the viral transfer , the
viral transfer the gene into
the nucleus but liposome has
not such intention.
• It is less efficient and its
game of chance.
• Doesn’t elicit immune
response .
Electroporation
• In this method we use short pulses of high voltage to
carry DNA across the cell membrane which makes a
shock to cause temporary formation of pores and thus
allow DNA molecule to pass.
• Electroporation has been used in vivo for many types of
tissue such as skin, muscle, lung gene delivery and tumor
treatment.
• Some of the draw back of electroporation can be
overcome by using of high voltage plasma discharge
DNA was efficiently delivered following very short
pulses, might damage tissue and affect genomic DNA
stability.
Gene gun
• In this method DNA is coated with gold particles and
loaded into a devise.
• Which is similar to gun and it generates force by
which it can penetrate into the cell.
• Gold or tungsten spherical particles are coated with
plasmid DNA and then accelerated to high speed by
pressurized gas to penetrate in to target tissue cells.
• Actually it is a modification technique called
“biolistic” originally developed for plant
transgenesis, but now used for in vitro and in vivo
gene delivery in to mammalian cell.
sonoporation
• Uses ultrasonic frequencies to deliver DNA into cell.
• Sonoporation is the transient permeation of cell
membranes assisted by ultrasound, typically in the
presence of gas micro bubbles.
• Sonoporation allows for the entry of genetic material
into cells.
• We use ultrasonic frequency to deliver DNA into cell.
• Which can disrupt the cell membrane and allow DNA
to move in to cell.
 Magnetofaction
• In this method the magnetic fields are used to concentrate
particles containing nucleic acid into target cell.
• In this way the magnetic force allows a very rapid
concentration of the entire applied vector dose onto cells.
• Magnetofaction has been adapted to all types of nucleic
acid , non viral transfection systems and viruses.
• Incase of in vivo , the therapeutic gene magnetic particle
complex is administered intravenously.
• Using strong high gradient external magnets, the complex
is captured and held at the target.
Nano particles
• The use of engineered inorganic and
organic nanoparticles is another non-viral approach
for gene delivery
• We use gold , silica and iron oxide and calcium
phosphate some of the benefit are storage stability,
low manufacturing cost and often time, low
immunogenicity.
Hybrid method
• There is another method other than viral and non viral
gene delivery system.
– i.e. hybrid gene delivery which is developed from
the combination of two or more technique.
• One of the hybrid method is virosomes which is
formed by the combination of liposomes with
activated HIV.
Vector Advantages Disadvantages
Retrovirus • High efficiency transduction of • Potential for insertional
appropriate target cells. mutagenesis.
• Long-term expression- integration • Requires dividing cells.
into chromosomal DNA). • Limited size of DNA insert.

Adenovirus • High transduction efficiency. • Transient expression.

• Broad range of target cells. • Immunogenicity.
• Does not require cell division. • Direct cytopathic effects of
• Low risk of insertional mutagenesis virus.
Adeno-associated • Does not require cell division. • Potential for insertional
virus (AAV) • Site specific integration. mutagenesis if integration
not site-specific.
• Limited size of DNA insert.
Non-viral • No infectious risk. • Low efficiency.
vectors(liposomes • Completely synthetic. • Limited target cell range.
, polymer based • No limitation on insert size. • Transient expression.
system)
PROBLEMS WITH GENE THERAPY

• Unwanted immune system reaction. Your body's immune system may see the
newly introduced viruses as intruders and attack them.

• Targeting the wrong cells.

• Possibility of causing a tumor.

• MULTIGENIC DISORDER = Heart disease ,high blood pressure , Alzheimer’s ,

arthritis and diabetes are hard to treat because we need to introduce more than
one gene.

• Organ damage
REFERENCES
http://www.webmd.boots.com/children/news/20150703/gene-therapy-cystic-fib
rosis

http://www.medscape.com/viewarticle/732741

https://www.youtube.com/watch?v=6mXQmSbc7ZE

http://www.shomusbiology.com/genetics1.html
wikipedia
• Group member Id no.
1. MINTESNOT FEKEDE…………...1059/13
2. TOFIK MUDESIR …….………....1164/13
3. ABRHAM TARIKU……………….. 712/13
4. MARUF MENSUR…………………..1105/13
5. REYAN AMERGA………………….1101/13
6. YONAS NEGASH…………………1012/13
7. ENDALE WONDIMU…………….1002/13
8. TOMAS ADUGNA………………...1178/13
9. ABUBEKER SHEMSU………………713/13