Content

 Introduction to Gene Therapy

 Importance of Gene Therapy
 Objective of the Project
 Theory: Basics of Genetics
 What is Gene Therapy?
 History of Gene Therapy
 Procedure of Gene Therapy
 Case Study: Sickle Cell Anemia
 Recent Advances in Gene Therapy
 Ethical Issues and Public Opinion
 Conclusion
 Bibliography

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Introduction
Gene therapy is a cutting-edge medical technique that aims to treat
or prevent diseases by directly modifying the genes within a person's
cells. Unlike conventional treatments that often manage symptoms,
gene therapy targets the underlying genetic cause of a disease. It
works by replacing, repairing, or removing faulty genes to restore
normal function.

Originally considered a
futuristic idea, gene therapy
has now become a reality
thanks to major advances in
genetic engineering,
biotechnology, and molecular
medicine. This project
explores the history,
mechanisms, procedures, and real-life applications of gene therapy,
as well as the challenges and ethical considerations surrounding its
use. With the potential to cure conditions once thought untreatable,
gene therapy represents a major leap forward in the field of medicine
and personalized healthcare.

Gene therapy is used in

Biotechnology
Medicine

Research

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Objective of the project:
To explore the structure and function of genes and their role in health
and disease.
To understand the principles, process, and purpose of gene therapy.
To examine the different types of gene therapy and how each functions
To study the major tools and techniques used in gene therapy,
including CRISPR and viral vectors.
To analyze real-life case studies and recent clinical successes.
To consider the ethical, social, and medical issues associated with
gene therapy and its future potenti

Theory
o What is DNA?
DNA (Deoxyribonucleic Acid) is the hereditary material in all living
organisms. It contains the genetic instructions
used in the growth, development, functioning,
and reproduction of all life forms. DNA is
shaped like a double helix and is made up of
four bases: adenine (A), thymine (T),
cytosine (C), and guanine (G).
These bases pair form genes, which are
instructions for making proteins. Proteins, in
turn, carry out most biological functions in
the body.

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o What is RNA?
RNA (Ribonucleic Acid) is a single-stranded molecule that helps
carry out the instructions in DNA. There are different types of RNA,
but the most important for gene therapy are:
 mRNA (messenger RNA): Carries genetic code from DNA to
ribosomes for protein synthesis.
 tRNA (transfer RNA) & rRNA (ribosomal RNA): Involved in the
process of protein production.
In gene therapy, synthetic or altered RNA can also be used to deliver
corrected genetic instructions.

What is Genetics?
Genetics is the study of genes, genetic variation, and heredity in
organisms. It explains how traits are passed from parents to offspring
and how mutations in genes can lead to diseases.
Some diseases are caused by:
 Single gene mutations
(e.g., sickle cell anemia)
 Chromosomal
abnormalities (e.g.,
Down syndrome)
 Multifactorial conditions
(e.g., diabetes)
Gene therapy attempts to
correct or replace faulty genes
responsible for such condition

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What Causes Genetic Disorders?
Genetic disorders occur when there are:
 Mutations in a single gene (monogenic disorders)
 Missing or extra chromosomes
 Complex interactions between multiple genes and the
environment
Examples:
 Cystic fibrosis – caused by mutation in the CFTR gene
 Hemophilia – mutation affects blood clotting genes
 Muscular dystrophy – mutations in the dystrophin gene
🧬 What is Gene Therapy?
Gene therapy involves introducing, altering, or removing genes
within a person's cells to treat or prevent disease. This can be done
by:
 Replacing a faulty gene with a
healthy one
 Inactivating a malfunctioning
gene
 Introducing a new or modified
gene to help treat a disease

Why is Gene Therapy Important?

 Treats the root cause of genetic diseases.
 Offers long-term or permanent solutions.
 Can reduce the burden of lifelong medication., Opens the door
to personalized medicine based on individual genetics.

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A Brief History of Gene Therapy
The idea of gene therapy emerged in the 1970s, but it remained
theoretical due to limited technology. A major milestone came in
1990, when a 4-year-old girl with Severe Combined
Immunodeficiency (SCID) became the first patient successfully
treated using gene therapy. Despite early challenges and safety
concerns, the field advanced
steadily
In 2012, Europe approved
Glybera, the first gene therapy
drug, for a rare disease called
Lipoprotein Lipase Deficiency
(LPLD).
Today, gene therapy is used or
studied for:
 Cancer (e.g., CAR-T cell
therapy)
 Rare genetic disorders (e.g., spinal muscular atrophy)
 Blood disorders (e.g., thalassemia, sickle cell anemia)
 Infectious diseases (e.g., mRNA-based approaches during
COVID-19)
Modern tools like CRISPR-Cas9, viral vectors, and nanoparticles
have made gene therapy more precise, safe, and effective than ever
before.

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Procedure
Gene therapy replaces or modifies faulty genes in a person’s cells to
treat genetic diseases. The process involves several key steps:
Step 1: Identification of the Faulty Gene
Doctors first identify the genetic disorder in
a patient by analyzing their DNA.
Techniques like gene mapping and DNA
sequencing help locate the defective gene
responsible for the disease.
Step 2: Creating a Functional Copy of the
Gene
Once the faulty gene is identified, scientists
create a healthy or normal copy of the same
gene in the lab. This gene will be used to
correct or replace the non-functioning gene in the patient.
Step 3: Gene Delivery Methods (Vectors)
Gene delivery is done using modified viruses called vectors that
safely insert the desired gene into the patient's cells. Common types
include:
1.Adenoviruses
o Cause mild illnesses (e.g., colds)
o Do not integrate into DNA → temporary effect
o Used for short-term treatments
2.Lentiviruses
o Can infect dividing and non-dividing cells
o Insert gene into host DNA → long-term expression
o Used for blood and inherited disorders

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3.Retroviruses
o Carry RNA and permanently integrate into DNA
o Infect dividing cells only
o Suitable for long-term treatment, but risk of insertion near
harmful genes
Step 4: Delivering the Gene into the Patient’s Cells
There are two primary delivery approaches:
 In vivo method: The therapeutic gene is injected directly into
the patient’s body, usually into the bloodstream or the specific
organ affected.
 Ex vivo method: The patient’s cells (like blood or bone marrow
cells) are first taken outside the body. The new gene is inserted
in the lab. These modified cells are then re-injected into the
patient
Step 5: Expression and Monitoring
Once inside the cells, the new gene begins to produce the required
functional protein that was missing or faulty due to the disease. The
patient is closely monitored over time to assess improvements and
detect any side effects.

Result
After gene therapy, the faulty gene is either replaced with a healthy
copy, inactivated, or supplemented with a new functional gene. As a
result, the corrected gene begins producing the right protein,
restoring the cell’s normal function. Patients often show marked
improvement in symptoms, and in many cases, gene therapy can
provide long-lasting or permanent relief. Continuous monitoring is
essential to ensure the treatment is safe and effective over time.

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 Case Study:
Gene Therapy
for Sickle Cell
Anemia

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🧬 Patient Background:
 Name (anonymized): Maria T.
 Age at treatment: 21 years
 Condition: Sickle Cell Anemia (SCA)
 Genetic Cause: A point mutation in the HBB gene,
which produces abnormal hemoglobin (Hemoglobin S)
 Symptoms: Severe joint and chest pain, fatigue, frequent
hospitalizations, anemia

⚕️Medical Problem:
Sickle cell anemia is an inherited blood disorder. Due to a single
base change in the HBB gene, red blood cells become
crescent-shaped, leading to:
 Poor oxygen delivery
 Blocked blood vessels
 Chronic pain
 Risk of organ damage and stroke
Maria had lived with frequent pain crises and had limited
mobility. Traditional treatments like blood transfusions and
hydroxyurea only managed symptoms.

🔬 Therapeutic Intervention – Gene Therapy:

 Goal: To produce healthy hemoglobin and prevent red
blood cells from sickling
 Method:
1. Maria’s hematopoietic (blood-forming) stem
cells were extracted from her bone marrow

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 2. A lentivirus was used to insert a modified gene that
produces functional fetal hemoglobin (HbF)
 HbF is not affected by the sickle mutation
3. After conditioning with chemotherapy to clear old
stem cells, the modified cells were infused back
into Maria

📈 Outcome:
 Within 6 months, her blood began showing a significant
rise in fetal hemoglobin
 She reported zero pain crises for the first time in years
 Blood tests confirmed healthier, round red blood cells
 No serious side effects occurred; her quality of life
dramatically improved

📌 Significance:
This case demonstrates how gene therapy can functionally
cure a lifelong disease. By reprogramming stem cells, gene
therapy allowed Maria’s body to bypass the genetic error
without needing a donor transplant.

🧠 What We Learn:
 Gene therapy is effective even for non-fatal but
debilitating diseases
 Lentiviral vectors are safe and efficient for treating
blood disorders
 The approach offers a long-term solution rather than
temporary relief

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Recent Advances in Gene Therapy

1.CRISPR-Cas9 Gene Editing

One of the most revolutionary tools in gene therapy is CRISPR-Cas9.
It allows scientists to precisely cut and modify specific genes,
correcting mutations at their source. Unlike earlier methods, CRISPR
is faster, more accurate, and relatively inexpensive.
✅ Example: CRISPR has shown promise in treating conditions like
sickle cell anemia, beta-thalassemia, and even some types of cancer.
2. FDA Approvals of Gene Therapy Drugs
Several gene therapies have received regulatory approval, marking a
shift from experimental to clinical use.
 Luxturna – treats a rare form of inherited blindness.
 Zolgensma – treats spinal muscular atrophy in infants.
 Roctavian – treats hemophilia A using an AAV (adeno-
associated virus) vector.
3. In Vivo vs Ex Vivo Techniques: Modern therapies have refined the
in vivo (inside the body) and ex vivo (outside the body, then
reintroduced) gene delivery approaches. Both methods have become
safer and more effective due to advancements in viral vectors and
nanotechnology.

4. Gene Therapy for Cancer: Gene therapy is being used to design

CAR-T cells, which are genetically modified immune cells that target
and destroy cancer cells. CAR-T therapy has been especially effective
in leukemia and lymphoma patients.

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thical Issues and Public Opinion
Gene therapy offers great hope but also raises important ethical and
social concerns that require careful consideration.
Germline vs. Somatic Therapy
Somatic therapy affects only the treated individual and is widely
accepted. Germline therapy
changes genes in reproductive
cells, passing changes to future
generations, raising concerns
about long-term safety and
consent.
Access and Equity
Gene therapies like Zolgensma are extremely expensive (over $2
million), limiting access mainly to wealthy patients and raising issues
of fairness and healthcare inequality worldwide.
Designer Babies
Advances like CRISPR have sparked fears of non-therapeutic genetic
enhancements (e.g., intelligence or appearance), which could
increase social inequality and reduce genetic diversity.
Religious and Cultural Views
Opinions vary widely; some support gene therapy for curing diseases,
while others see it as “playing God,” especially concerning germline
editing.
Informed Consent and Safety
Patients must be fully informed about risks and benefits and
participate voluntarily, with research conducted under strict ethical
guidelines.

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Conclusion
 Gene therapy is a revolutionary advancement in medical
science and biotechnology.
 It treats or potentially cures genetic disorders by correcting
defects in DNA.
 Gene therapy has progressed from research to successful
clinical use.
 It shows promising results in diseases like spinal muscular
atrophy, certain cancers, and inherited blindness.
 This project covered the principles, types, applications, case
studies, and recent developments of gene therapy.
 Techniques such as CRISPR-Cas9 and viral vectors enable
precise gene editing.
Challenges include:
o High treatment costs and unequal access.
o Ethical concerns about genetic enhancement and long-
term safety.
 These issues require careful regulation and public discussion.
 Gene therapy must prioritize human well-being, fairness, and
safety.
 With ethical guidance and equitable access, gene therapy can
transform healthcare.
 It offers hope to millions suffering from genetic diseases.
 Continued research may redefine disease treatment and
prevention in the future.

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Whole document’s summary
(for student refrence)
🔹 What is Gene Therapy?

 A medical technique that treats or prevents disease by modifying a person’s

genes.
 Targets the root cause of genetic disorders, unlike regular treatments that
just reduce symptoms.

🔹 Importance

 Can cure diseases like SCID, sickle cell anemia, hemophilia.

 Offers permanent solutions by correcting faulty genes.
 Reduces the need for lifelong medication.

🔹 Objective of the Project

 Understand how gene therapy works.

 Study its types, tools (like CRISPR, viral vectors), applications, and ethical
concerns.

🔹 Theory Highlights

 DNA: Genetic blueprint, made of A, T, C, G bases.

 RNA: Helps in protein formation; mRNA is key in gene therapy.
 Genetics: Study of heredity; gene mutations can cause disorders.

🔹 Causes of Genetic Disorders

 Single gene mutations (e.g., cystic fibrosis)

 Chromosome abnormalities (e.g., Down syndrome)
 Multifactorial (genes + environment)

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🔹 Types of Gene Therapy

 Gene replacement (replace faulty gene)

 Gene inactivation (switch off bad gene)
 Gene insertion (add new functional gene)

🔹 Brief History

 First trial in 1990 (SCID patient).

 Glybera (2012) was first approved therapy in Europe.
 Now used in cancer, rare diseases, and more.

🔹 Procedure (Steps)

1. Identify faulty gene

2. Make correct gene in lab
3. Choose a vector (like virus – adenovirus, lentivirus, retrovirus)
4. Deliver gene (in vivo or ex vivo)
5. Monitor patient for response and safety

🔹 Case Study – Sickle Cell Anemia

 Patient “Maria” had severe symptoms.

 Her stem cells were edited using a lentivirus to make healthy hemoglobin.
 Result: No more pain crises, improved life quality.

🔹 Recent Advances

 CRISPR-Cas9: Edits genes with precision.

 Approved therapies: Zolgensma (SMA), Luxturna (blindness), Roctavian
(hemophilia).
 CAR-T therapy: Gene-edited immune cells for cancer treatment.

🔹 Success Stories

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  Sickle Cell: 90% effectiveness with CRISPR.
 SMA (Zolgensma): Saved infants’ lives.
 Blindness (Luxturna): Restored partial vision.
 Cancer (CAR-T): Over 80% remission in leukemia.

🔹 Ethical Concerns

 Germline editing affects future generations—raises safety & consent issues.

 High treatment costs limit access.
 Designer babies debate—modifying traits like looks or intelligence.
 Need for strict regulation, public discussion, and informed consent.

🔹 Conclusion

 Gene therapy is transforming medicine.

 It corrects genetic errors at the DNA level.
 Still faces ethical, financial, and accessibility challenges.
 With responsible use, it offers hope for millions.

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