Types of Gene
Therapy
Outline
Introduction
✓ Gene therapy is a therapeutic approach aimed at treating or preventing diseases by
modifying the genetic material of cells.
✓ This typically involves introducing, altering, or removing specific genes within an
individual's cells to correct a genetic defect or to provide a therapeutic effect.
Introduction
✓ Gene therapy holds immense importance in modern medicine due to its potential
to revolutionize the treatment of genetic disorders, infectious diseases, and various
types of cancers.
❖ Specific transcription factors in
cancer cells are able to induce
therapeutic gene expression.
❖ On the other hand, expression of the
therapeutic gene is not obtained in
normal cells because they lack the
specific transcription factors
Fundamentals
of Genetics
Genes and DNA
✓ Genes: Fundamental units of heredity, composed of DNA, carrying
instructions for the development, functioning, growth, and
reproduction of organisms.
✓ DNA (Deoxyribonucleic Acid): The molecule that encodes genetic
instructions. It is the blueprint of life, carrying genetic information
in all living organisms.
Genes and DNA
✓ Transcription: Process where DNA is used as a template to produce mRNA (messenger RNA).
✓ Translation: Process where mRNA is decoded by ribosomes to produce specific proteins, essential
for various cellular functions.
Genetic Diseases
✓ Genetic disorders are conditions caused by abnormalities
in an individual's genetic makeup, usually inherited from
one or both parents.
✓ Genetic disorders can affect various aspects of health,
development, and quality of life.
Types of Genetic Disorders
Examples: Down syndrome (trisomy 21), Turner
syndrome (monosomyX), Klinefelter syndrome
(XXY).
Chromosomal
Disorders:
Cause: Abnormalities in the number or structure
of chromosomes.
Examples: Cystic fibrosis, sickle cell anemia,
Huntington's disease.
Single-Gene
Disorders:
Cause: Mutations or alterations in a single gene.
Examples: Diabetes, heart disease, certain types
of cancer.
Multifactorial
Disorders:
Cause: Interaction between genetic
predisposition and environmental factors.
Types of Gene Therapy
Germ Ex vivo
Line vs. In
Gene vivo
Therapy Therapy
Somatic
Gene
Therapy
Germ Line Gene Therapy
✓ Definition: Germ line gene therapy involves modifying the DNA of germ cells (sperm or eggs) to
introduce heritable genetic changes that can be passed on to future generations.
✓ Purpose: To address genetic disorders at their root cause by correcting or preventing disease-
causing mutations in the germ line.
How Germ Line Gene Therapy Works
✓ Process: Germline gene therapy involves introducing desired genetic modifications into germ cells
using various techniques, such as viral vectors or genome editing tools like CRISPR-Cas9.
✓ Target Cells: Germline gene therapy targets sperm or eggs to ensure that the genetic changes are
passed on to offspring.
CRISPR-Cas9
✓ Definition: CRISPR-Cas9 is a powerful genome editing tool derived from a bacterial defense
system that allows precise modification of DNA sequences.
✓ Guide RNA (gRNA): Designed to target a specific DNA sequence complementary to the target
region.
✓ DNA Cleavage: Cas9 enzyme cuts the DNA at the target site, inducing a double-strand break.
✓ Repair Mechanisms: Cellular repair mechanisms such as non-homologous end joining (NHEJ) or
homology-directed repair (HDR) can then be utilized to introduce desired genetic modifications.
CRISPR-Cas9
Somatic Gene Therapy
✓ Definition: Somatic gene therapy involves the introduction of therapeutic genes into the cells of
an individual's body to treat or alleviate the symptoms of a specific disease.
✓ Purpose: Unlike germ-line gene therapy, somatic gene therapy does not affect the individual's
germ cells and thus does not result in heritable genetic changes.
Somatic Gene Therapy Vs Germline Therapy
Mechanism of Action
✓ Delivery to Target Cells: Therapeutic genes are introduced into target cells using viral or non-viral
vectors.
✓ Expression of Therapeutic Proteins: Once inside the cell, the therapeutic genes are expressed,
leading to the production of therapeutic proteins or RNA molecules.
✓ Correction of Disease: The therapeutic proteins or RNA molecules act to correct the underlying
molecular defects or modulate cellular functions associated with the disease.
Mechanism of Action
Vectors
Ex vivo vs. In
vivo Therapy
Ex vivo Gene Therapy
✓ Definition: In ex vivo gene therapy, cells are isolated from the patient, genetically modified
outside the body, and then reintroduced into the patient.
Process:
Genetic Modification: The Reinfusion: The
Isolation: Cells, often Expansion: Modified cells
therapeutic gene is genetically modified cells
stem cells or immune introduced into the are cultured and are reintroduced into the
cells, are extracted from expanded to achieve the
isolated cells using viral patient's body, where they
the patient. or non-viral vectors. desired number. exert therapeutic effects
In vivo Gene Therapy
✓ Definition: In in vivo gene therapy, therapeutic genes are directly delivered into the patient's
body, targeting specific tissues or organs.
Process:
Therapeutic Effect: The
Delivery: Therapeutic genes Expression: Once inside the
expressed proteins or RNA
are delivered directly into the Targeting: The delivery system body, the therapeutic genes are
molecules correct the
patient's body using viral targets specific tissues or cells expressed, leading to the underlying molecular defects or
vectors, non-viral vectors, or affected by the disease. production of therapeutic
modulate cellular functions
gene editing technologies. proteins or RNA molecules.
associated with the disease.
Ex vivo vs. In vivo Therapy
Gene Delivery Systems
Viral Vector-
based Non-viral
Delivery Vector-based
Delivery
Viral Vector-based Delivery
✓ Definition: Viral vectors are modified viruses
engineered to carry therapeutic genes into target
cells.
✓ Properties: Viral vectors retain the ability to
infect cells and deliver genetic material but are
modified to remove their ability to cause disease.
Types of Viral Vectors
Adeno-
Retroviral Adenoviral Associated
Vectors: Vectors: Viral (AAV)
Vectors:
Used for stable,
long-term gene Derived from
Efficiently infect a adeno-associated
Derived from expression, wide range of Highly efficient
retroviruses, Derived from viruses, which and safe, with low
making them adenoviruses, dividing and non-
which integrate can establish immunogenicity,
suitable for which do not dividing cells,
their genetic treating diseases long-term making them
integrate into the making them episomal gene
material into the requiring suitable for suitable for in vivo
host cell's host genome and expression gene therapy
continuous remain episomal. transient gene
genome. without applications.
protein expression.
production. integration.
Non-viral Vector-based Delivery
✓ Definition: Non-viral vectors are synthetic or naturally derived carriers used to deliver
therapeutic genes into target cells without the use of viral components.
✓ Properties: Non-viral vectors are typically safer, less immunogenic, and easier to manufacture
compared to viral vectors.
Types of Non-viral Vectors
Nucleic Acid-based
Lipid-based Vectors: Polymeric Vectors: Vectors:
RNA-based Vectors:
Synthetic RNA
Polymers: Synthetic or Polyplexes:
Liposomes: Synthetic Lipid nanoparticles: naturally derived Complexes formed Plasmid DNA: Circular molecules, such as
lipid vesicles that Lipid-based DNA molecules messenger RNA
polymers that between polymers and
encapsulate nanoparticles that condense therapeutic therapeutic genes, containing therapeutic (mRNA) or small
therapeutic genes and protect therapeutic genes that can be interfering RNA
genes into facilitating their (siRNA), used for
facilitate their delivery genes and enhance nanoparticles for internalization into delivered directly into
into cells. their cellular uptake. delivery. cells. target cells. transient gene
expression or gene
silencing.
Non-viral Vector-based Delivery
Lipid-
based
Vectors:
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