Biology

Project
Report
On

“Gene Therapy”
Session: 2020-21”

Submitted to:- Submitted by

Mr. Raj Kamal Sarita

Lecturer in Biology Roll No.
Class XII (Medical)
 Sant Gyaneshwar Model
School Alipur
Delhi
CERTIFICATE

Certified that the investigatory project entitled “Gene Therapy” was carried

out in Biology by Sarita, a student of XII (Medical), Sant Gyaneshwar Model

School Alipur Delhi as a partial fulfilment of the practical work conducted by the

Central Board of Secondary Education, New Delhi.

Mr. Raj Kamal

Lecturer in Biology
 ACKNOWLEDGEMENT

I take this opportunity to thank and express my sincere gratitude to

Mr. Raj Kamal, Lecturer in Biology who guided me for this project and without
whose efforts; this project would not have been completed. I put my sincere efforts
to make this project interesting. I fully consulted all the available books on this
subject and I am thankful to esteemed authors.

In the end, I am thankful to all those who took interest in the successful
completion of the project.

Sarita
XII (Medical)
Roll No. ................................
 CONTENT

1. Introduction

2. Types of Gene Therapy

3. Approaches in GENE Therapy

4. Ex-Vivo Gene Therapy

5. In Vivo Gene Therapy

6. Vectors in Gene Therapy

7. Method of Gene Delivery

8. Chemical Methods

9. Other types of Gene Therapy

10. Advantage & Disadvantage

11. Ethical Issues

12. Conclusion
 INTRODUCTION

Gene therapy is the introduction of genes into existing cells to prevent or cure a wide
range of disease.

It is a technique for correcting defective genes responsible for disease development.

The first approved gene therapy experiment ocured on September 14, 1990 in US, when
Ashanti DeSilva was treated for ADA-SCID.

1
 TYPES OF GENE THERAPY

SOMATIC CELL GENE THERAPY GERM LINE GENE THERAPY

 Therapeutic Genes transferred into the  Therapeutic genes transferred into

somatic cells. the germ cells.

 Eg. Introduction of genes into bone  Eg. Genes introduced into eggs and
marrow cells, blood cells, skin cells etc. sperms.

 Will not be inherited later generations.  It is heritable and passed on to later

generations.

 At present all researches directed to  For safety, ethical and technical

correct genetic defects in somatic cells. reasons, it is not being attempted at
present.

2
 APPROACHES IN GENE THERAPY

 In vivo Gene Therapy:

Direct delivery for genes into the cells of particular tissue in the body.

 Ex vivo Gene Therapy:-

Transfer of genes to cultured cells and reinsertion.

3
EX VIVO GENETHERAPY

4
EXAMPLE OF EX VIVO GENE THERAPY

 1ST Gene Therapy – to correct deficiency of enzyme, Adenosine deaminase

(ADA).
 Performed on a 4yr old girl Ashanthi DeSilva.
 Was suffering from SCID-Severe Combined Immunodeficiency.
 Caused due to defect in gene coding for ADA.
 Deoxy adenosine accumulate and destroys T lymphocytes.
 Disrupts immunity, suffer from Infectious diseases and die at young age.

5
 IN VIVO GENE THERAPY

 Direct delivery of therapeutic gene into target cell into patients body.
 Carried out by viral or non viral vector systems.
 It can be the only possible option in patients where individual cells cannot be
cultured in vitro in sufficient numbers (e.g. brain cells).
 In vivo gene transfer is necessary when cultured cells cannot be re-implanted in
patients effectively.

EXAMPLE OF IN VIVO GENE THERAPY

Therapy for Cystic Fibrosis:

 In patients with cystic fibrosis, a protein called cystic fibrosis transmembrane

regulator (CFTR) is absent due to a gene defect.

6
 In the absence of CFTR chloride ions concentrate within the cells and it draws
water from surrounding.
 This leads to the accumulation of sticky mucous in respiratory tract and lungs.
 Treated by in vivo replacement of defective gene by adenovirus vector.

7
 VECTORS IN GENE THERAPY

 To transfer the desired gene into a target cell, a carrier is required. Such vehicles
of gene delivery are known as vectors.
 2 main classes
o Viral vectors
o Non viral vectors

Viral Vectors

1. Retrovirus Vector System

 The recombinant retroviruses have the ability

to integrate into the host genome in a stable
fashion.
 Can carry a DNA of size – less than 3.4kb
 Replication defective virus particles
 Target cell – dividing

2. Adeno Virus Vector System

8
  Adeno virus with a DNA genome good vectors.
 Target – non-dividing human cell.
 Eg. Common cold adenovirus

3. Adeno Associated Virus Vector

 It is a human virus that can integrate into chromosome 19.

 It is a single stranded, non pathogenic small DNA virus.
 AAV enters host cell, becomes double stranded and gets integrated into
chromosome.

4. Herpex Simplex Virus Vector

 Viruses which have natural tendency to infect a particular type of cell.

 They infect and persist in nervous cells.

NON VIRAL VECTOR SYSTEM

1. Pure DNA Construct

 Direct introduction of pure DNA construct into target tissue.

 Efficiency of DNA uptake y cells and expression rather low.
 Consequently, large quantities of DNA have to be injected periodically.

2. Lipoplexes

 Lipid DNA complexes; DNA construct surrounded by artificial lipid layer.

 Most of it gets degraded by lysosomes.

3. DNA Molecular Conjugates

9
  Commonly used synthetic conjugate is poly – L – lysine bound to specific target
cell receptor.
 Therapeutic DNA is then made to combine with the conjugate to form a complex.
 It avoids lysosomal breakdown of DNA.

4. Human Artificial Chromosome

 Can carry a large DNA ie, with one or more therapeutic genes with regulatory
elements.

METHODS OF GENE DELIVRY

10
PHYSICAL METHODS

Gene Gun

 Employs a high-pressure delivery system to shoot

tissue with gold or tungsten particles that are
coated with DNA

Microinjection

 Process of using a glass micropipette to insert

microscopic substances into a single living
cell.
 Normally performed under a specialized
optical microscope setup called a
micromanipulator.

CHEMICAL METHODS

11
Using Detergent Mixtures

 Certain charges chemical compounds like Calcium phosphates are mixed with
functional cDNA of desired function.
 The micxture is introduced near the vicinity of recipient cells.
 The chemical disturbs the cell membrance, widens the pore size and allows cDNA
to pass through the cell.

Lipofection

 It is a technique used to inject genetic materials into a cell by means of liposomes.

 Liposomes are artificial phospholipid vesicles used to deliver a variety of
molecules including DNA into the cells.

OTHER TYPES OF GENE THERAPY

12
Gene Augmentation Therapy

 Most common form of gene therapy

 Foreign gene replaces missing or defective gene.
 Eg. Replacement of defective p53 gene by a normal one in liver cancer.

Gene Inhibition Therapy

 Done to block the overproduction of some proteins.

 2 types – Antigene and Antisense therapy
o Antigene – blocks transcription using antigene oligonucleotide.
o Antisense – blocks transalation using antisense oligonucleotide.

ADVANTAGES

13
 Gene therapy has the potential to eliminate and prevnt hereditary diseases sch as
cystic fibrosis, ADA-SSID etc.
 It is a possible cure for heart disease, AIDS and canser.
 It gives someone born with a genetic disease a chance to life.
 It can be used to eradicate diseases from the future generations.

DISADVANTAGES

 Long lasting therapy is not achieved by gene therapy; Due to rapid dividing of
cells benefits of gene therapy is short lived.
 Immune response to the transferred gene stimulates a potential risk to gene
therapy.
 Viruses used as vectors for gene transfer may cause toxicity, immune responses,
and inflammatory reactions in the host.
 Disorder caused by defects in multiple genes cannot be treated effectively using
gene therapy.

ETHICAL ISSUES

14
 Who will have access to therapy?
 Is it interfering with God’s plan?
 Should people be allowed to use gene therapy to enhance basic human trait such as
height, intelligence etc.?
 It is alright to use the therapy in the prenatal stage of development in babies?

CONCLUSION

15
  Theoretically, gene therapy is the permanen solution for genetic diseases.
 But it has several complexities. Al its current stage, it is not accessibl to most
people due to its huge cost.
 A breakthrough may come anytime and day may come when almost every disease
will have a gene therapy.
 Gene therapy have the potential to revolutionize the practice of medicine.

RECENT DEVELOPMENT

 In a new gene the therapy method developed by University of

Florida in Jan 2012, researchers found treatment for a common form of blindness
(X-linked retinitis pigmentosa) that strikes both youngsters and adults.
 A gene therapy called NLX-P101 dramatically reduces movement impairment in
Parkinson’s patients, according to results of a Phase 2 study published on March,
2011 in the journal Lancent Neurology.

16