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Biology Investigatory

Gene Therapy

Name: Ragib Javed


Year: 2013-14
This is to certify that OV Shashank a student of
RN Podar School, of class XII- D, Roll
No: , has completed his full semester
project in the fulfilment of curriculum All India
Senior Secondary Examination
The project work entitled Gene Therapy, is the
original work done by him during his above
full semester project.

_____________________ ___________________
Internal Examiner Principal

External Examiner School Stamp


I take this opportunity to express my sincere gratitude
to the honourable Principal Mrs Avnita Bir of RN
Podar School for her deep interest and guidance
provided to me during the course.
I am most grateful to our Biology teacher Mrs
Padmavathi for her great help in the completion of
this project.

Students Signature

Table of Contents

Gene Therapy
Types of gene therapy
Isolation of gene
Gene Targeting
Gene Delivery
Case Study Cystic Fibrosis
The Disease
Is it a good Target
Choosing Vectors
Ethical Issues
Recent Upcoming


The term disease broadly refers to any condition that impairs normal
function, and is therefore associated with dysfunction of normal
homeostasis. When the functioning of one or more organs or systems of
the body is adversely affected, characterised by various signs and
symptoms, we say that we are not healthy, i.e., we have a disease.

Health can be defined as a state of complete physical, mental and social

well-being. When people are healthy, they are more efficient at work.
This increases productivity and brings economic prosperity. Health also
increases longevity of people and reduces infant and maternal mortality.

In humans, disease is often used more broadly to refer to any condition

that causes pain, dysfunction, distress, social problems, or death to the
person afflicted, or similar problems for those in contact with the person.
In this broader sense, it sometimes
includes injuries, disabilities, disorders, syndromes, infections,
isolated symptoms, deviant behaviours, and atypical variations of
structure and function, while in other contexts and for other purposes
these may be considered distinguishable categories.

Based on the cause diseases can be broadly classified as:

These are diseases caused due to invasion of a
foreign parasitic organism. They are temporary
because the immune system of organisms can fight
such pathogens (disease causing organisms) to a
certain extent hence helping in prevention of the
disease. The immune system can also be aided
with the use of several drugs. Apart from easy treatment they can also
be easily prevented.

Lifestyle Diseases
Lifestyle diseases (also sometimes called diseases of
longevity or diseases of civilization interchangeably) are diseases that

appear to increase in frequency as countries become more industrialized
and people live longer. Diet and lifestyle are major factors thought to
influence susceptibility to many diseases. Drug abuse, tobacco smoking,
and alcohol drinking, as well as a lack of exercise may also increase the
risk of developing certain diseases, especially later in life. These
diseases can be prevented completely by living a healthy lifestyle.

Certain diseases, such as diabetes, dental caries and asthma, appear at

greater rates in young populations living in the "western" way; their
increased incidence is not related to age, so the terms cannot accurately
be used interchangeably for all diseases.

Genetic Disorders
A genetic disorder is an illness caused by one or more abnormalities in
the genome, especially a condition that is present from birth
(congenital). They are medical disorders related to gene mutation.

Genetic disorders are heritable, and are passed down from the parents'
genes. Other defects may be caused by new mutations or changes to
the DNA. In such cases, the defect will only be
heritable if it occurs in the germ line. The
same disease, such as some forms of cancer,
may be caused by an inherited genetic
condition in some people, by new mutations in
other people, and by non-genetic causes in still
other people.

These diseases are totally random and difficult to prevent as they are not
caused by external agents. Also as their root cause lies in the genome of
the organism their cure was thought to be impossible until the
breakthrough research unlocking the secrets of DNA leading to the
development of biotechnology and hence gene therapy.

Gene Therapy
We can think of a medical
condition or illness as a
"broken window." Many
medical conditions result
from flaws, or mutations, in
one or more of a person's
genes. Mutations cause
the protein encoded by
that gene to malfunction.
When a protein
malfunctions, cells that
rely on that protein's
function can't behave
normally, causing problems for whole tissues or organs.
Medical conditions related to gene mutations are called
genetic disorders.

So, if a flawed gene caused our "broken window," can we

"fix" it? What are our options?

1. Stay silent: ignore the genetic disorder and nothing

gets fixed.

2. Try to treat the disorder with drugs or other

approaches: depending on the disorder, treatment
may or may not be a good long-term solution.

3. Put in a normal, functioning copy of the gene: if you

can do this, it may solve the problem!

If it is successful, gene therapy provides a way to fix a

problem at its source. Adding a corrected copy of the gene
may help the affected cells, tissues and organs work
properly. Gene therapy differs from traditional drug-based

approaches, which may treat the problem, but which do
not repair the underlying genetic flaw.

Gene therapy is the therapeutic delivery of nucleic acid polymers into a

patient's cells as a drug to treat disease. The first therapeutic use of
gene transfer as well as the first direct insertion of human DNA into the
nuclear genome was performed by French Anderson in a trial starting in
September 1990.

Targets for Gene Therapy

But now a question arises, which disorders or diseases
can we target using gene therapy? Many disorders or
medical conditions might be treated using gene therapy,
but others may not be suitable for this approach. For a
disease to be targeted by gene therapy it must satisfy the
following conditions:

1. The condition must result from mutations in one or

more genes.

2. To treat a genetic flaw, the knowledge of which

gene(s) to pursue is absolutely necessary. A DNA copy
of that gene available in the laboratory. The best
candidates for gene therapy are the so-called "single-
gene" disorders - which are caused by mutations in
only one gene.

3. To design the best possible approach, knowledge

about how the gene factors into the disorder is
required. For example:

Which tissues are affected?

What role does the protein encoded by the gene

play within the cells of that tissue?

Exactly how do mutations in the gene affect the

protein's function?

4. Adding a normal copy of the gene should fix the
problem in the affected tissue. This may seem like
obvious, but it's not. What if the mutated gene
encodes a protein that prevents the normal protein
from doing its job? Mutated genes that function this
way are called dominant negative and adding back
the normal protein won't fix the problem.

5. The gene delivery to cells of the affected tissue must

be possible. It depends on:

How accessible is the tissue? Is it fairly easy

(skin, blood or lungs), or more difficult to reach
(internal organs)?

What is the best mode of delivery?

The techniques of biotechnology have made it possible to

isolate the required gene in the laboratory and also deliver
the gene.

Types of Gene

Cell types
Gene therapy may be classified into two types:

In somatic cell gene therapy (SCGT), the therapeutic
genes are transferred into any cell other than
a gamete, germ cell, gametocyte or undifferentiated stem
cell. Any such modifications affect the individual patient
only, and are not inherited by offspring. Somatic gene
therapy represents mainstream basic and clinical
research, in which therapeutic DNA (either integrated in
the genome or as an external episome or plasmid) is used
to treat disease.

Over 600 clinical trials utilizing SCGT are underway in the

US. Most focus on severe genetic disorders,
including immunodeficiencies, haemophilia, thalassaemia
and cystic fibrosis. Such single gene disorders are good
candidates for somatic cell therapy. The complete
correction of a genetic disorder or the replacement of
multiple genes is not yet possible. Only a few of the trials
are in the advanced stages.

In germline gene therapy (GGT), germ
cells (sperm or eggs) are modified by the introduction of
functional genes into their genomes. Modifying a germ cell
causes all the organism's cells to contain the modified
gene. The change is therefore heritable and passed on to
later generations. Australia, Canada, Germany, Israel,

Switzerland and the Netherlands prohibit GGT for
application in human beings, for technical and ethical
reasons, including insufficient knowledge about possible
risks to future generations and higher risks versus
SCGT. The US has no federal controls specifically
addressing human genetic modification (beyond FDA
regulations for therapies in general).

Isolation of Gene of Interest

The first step is to find and isolate the gene that will be
inserted into the genetically modified organism. Finding
the right gene to insert usually draws on years of
scientific research into the identity and function of useful
genes. Once that is known the DNA needs to be cut at
specific locations to isolate the gene of interest. This can
be done by using restriction enzymes also known as
molecular scissors which cut DNA at specific sites
containing palindromic DNA sequences. But in order to cut
the DNA with restriction enzymes, it needs to be in pure
form, free from other macro-molecules.
Until the early 1970s DNA was the most difficult
cellular molecule for the biochemist to analyse.
Enormously long and chemically monotonous, the string of
nucleotides that forms the genetic material of an
organism could be examined only indirectly,
by protein or RNA sequencing or by genetic analysis.
Today the situation has changed entirely. From being the
most difficult macromolecule of the cell to analyse, DNA
has become the easiest.

Isolation of DNA
Since the DNA is enclosed within the membranes, we have
to break the cell open to release DNA
along with other macromolecules such
as RNA, proteins, polysaccharides and
also lipids. This can be achieved by
treating the bacterial cells/plant or
animal tissue with enzymes such as
lysozyme (bacteria), cellulase (plant
cells), chitinase (fungus). Genes are
located on long molecules of DNA
intertwined with proteins such as histones. The RNA can
be removed by treatment with ribonuclease whereas
proteins can be removed by treatment with protease.
Other molecules can be removed by appropriate
treatments and purified DNA ultimately precipitates out
after the addition of chilled ethanol. This can be seen as
collection of fine threads in the suspension.

Cutting of DNA
Restriction enzyme digestions are
performed by incubating purified
DNA molecules with the
restriction enzyme, at the optimal
conditions for that specific
enzyme. The cutting of DNA by
restriction endonucleases results
in the fragments of DNA. These fragments can be
separated by a technique known as gel electrophoresis.
Since DNA fragments are negatively charged molecules
they can be separated by forcing them to move towards
the anode under an electric field through a
medium/matrix. The separated bands of DNA are analysed

for the required gene and then it is cut out from the
agarose gel and extracted from the gel piece. This step is
known as elution.

Multiplication of Gene (PCR)

PCR or polymerase chain reaction is then used to create
multiple copies of the gene of interest. In this reaction,
multiple copies of the gene (or DNA) of interest is
synthesised in vitro using two sets of primers (small
chemically synthesised oligonucleotides that are
complementary to the regions of DNA) and the enzyme
DNA polymerase. The enzyme extends the primers using
the nucleotides provided in the reaction and the genomic
DNA as template. If the process of replication of DNA is
repeated many times, the segment of DNA can be
amplified to approximately billion times, i.e., 1 billion
copies are made.

PCR is a wonderful technology for amplifying DNA. It

allows you to take a specific region of DNA on the
chromosome and through the use of primers, copy back
and forth, only a particular desired segment, making two,
then four, then eight, then sixteen, and so on, up to
millions of copies. It is possible to start from the DNA
segment of a single cell and produce enough of it for use
in DNA typing or fingerprinting.
PCR relies on the ability of DNA-copying enzymes to
remain stable at high temperatures. The process was
sparked by a high temperature bacterium (Thermus
aquaticus) inhabiting the hot springs of Yellowstone
National Park.

Gene Targeting
Gene targeting (also, replacement strategy based on
homologous recombination) is a genetic technique that
uses homologous recombination to change
an endogenous gene. The method can be used to delete a
gene, remove exons, add a gene, and introduce point
mutations. Gene targeting can be permanent or
conditional. Conditions can be a specific time
during development / life of the organism or limitation to a
specific tissue.

Gene delivery is one of the biggest challenges in

the field of gene therapy.

Gene Delivery includes:

1. TARGETING the right cells.

2. ACTIVATING the gene. A gene's journey is not over when

it enters the cell. It must go to the cell's nucleus and be
"turned on," meaning that its transcription and translation
are activated to produce the protein product encoded by
the gene. For gene delivery to be successful, the protein
that is produced must function properly.

3. INTEGRATING the gene in the cells. The gene must stay

put and continue working in the target cells. If so, it must
be ensured that the gene integrates into, or becomes part
of the host cell's genetic material, or that the gene finds
another way to survive in the nucleus without being

4. AVOIDING harmful side effects. Anytime an unfamiliar

biological substance is introduced into the body, there is a
risk that it will be toxic or that the body will mount an

immune response against it. If the body develops immunity
against a specific gene delivery vehicle, future rounds of
the therapy will be ineffective. For gene delivery to be
successful, foreign DNA must survive long enough in the
host cell to integrate into its genome. 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 .Vectors utilized as
the method for gene delivery can be divided into two
categories, non-viral and viral.

Choosing the Best Vector

There is no "perfect vector" that can treat every disorder.
Like any type of medical treatment, a gene therapy vector
must be customized to address the unique features of the
disorder. We have learnt the lesson, of transferring genes
into plants and animals from bacteria and viruses, which
have known this for ages how to deliver genes to
transform eukaryotic cells and force them to do what the
bacteria or viruses want.

Gene therapy utilizes the delivery of DNA into cells,

which can be accomplished by several methods,
summarized below. The two major classes of methods are
those that use recombinant viruses (sometimes called
biological nanoparticles or viral vectors) and those that
use naked DNA or DNA complexes (non-viral methods).

Part of the challenge in gene therapy is choosing the most

suitable vector for treating the disorder. Some vectors
commonly used are:


Usually when we think of viruses, we think of them

causing diseases such as the common cold, the flu, and

HIV/AIDS. When faced with the problem of gene delivery,
scientists looked to viruses. Why reinvent the wheel if
there's a perfectly good one out there? If we can modify
viruses to deliver genes without making people sick, we
may have a good set of gene therapy tools.

All viruses bind to their hosts and introduce their genetic

material into the host cell as part of their replication
cycle. This genetic material contains basic 'instructions'
of how to produce more copies of these viruses, hacking
the body's normal production machinery to serve the
needs of the virus.

The host cell will carry out these instructions and produce
additional copies of the virus, leading to more and more
cells becoming infected. Some types of viruses insert
their genome into the host's cytoplasm, but do not
actually enter the cell. Others penetrate the cell
membrane disguised as protein molecules and enter the

General advantages of viral vectors:

-They're very good at targeting and

entering cells.

-Some viral vectors might be

engineered to target specific
types of cells.

-They can be modified so that they can't replicate and

destroy the cell.

General drawbacks of viral vectors:

A virus can't "expand" to fit a piece of genetic material

larger than it is naturally built to carry. Therefore, some
genes may be too big to fit into a certain type of virus.

Viruses can cause immune responses in patients,
resulting in two potential outcomes:

Patients may get sick.

A patient's immunity to a virus may prevent him from

responding to repeated treatments.

However, modern viral vectors have been engineered

without most of the proteins that would cause an immune

Non-Viral Vectors

Although viruses can effectively deliver genetic material

into a patient's cells, they do have some limitations. It is
sometimes more efficient to deliver a gene using a non-
viral vector, which has fewer size constraints and which
won't generate an immune response.

Non-viral vectors are typically circular DNA molecules,

also known as plasmids. In nature, bacteria use plasmids
to transfer genes from cell to cell.

Scientists use bacteria and plasmids to easily and

efficiently store and replicate genes of interest from any

Vectors used at present, are engineered in such a way that

they help easy linking of foreign DNA and selection of
recombinants from non-recombinants.

These are not the only way to introduce alien DNA into host cells. In a
method known as micro-injection, recombinant DNA is
directly injected into the nucleus of an animal cell. In
another method, suitable for plants, cells are bombarded
with high velocity micro-particles of gold or tungsten

coated with DNA in a method known as biolistics or gene

Delivery to specific tissues

Delivering genes to specific tissues within a patient's

body can be very difficult. Delivering genes into a group of
cells in a patient's body can be done in one of two ways.

The first way is to inject the vector into the body and
specifically target affected cells. This is called an in vivo
approach. The second way, called ex vivo, is to deliver the
gene to cells while they're outside the body by:

Isolating the desired cells from the body.

Culturing the cells in a Petri dish in the laboratory.

Delivering the genes to the cells (using one of the

vector options described on this page), activating them,
and making sure that the cells integrate them properly.

Case Study
Cystic Fibrosis

The Disease A Genetic Disorder

Cystic fibrosis (CF), also known as

mucoviscidosis, is an autosomal
recessive genetic disorder that affects
most critically the lungs, and also the
pancreas, liver, and intestine. It is
characterized by abnormal transport of
chloride and sodium across an epithelium,
leading to thick, viscous secretions,
preventing the cilia from clearing debris
which cause symptoms such as coughing,
poor digestion and increased vulnerability
to infection.

CF is caused by a mutation in the gene for

the protein cystic fibrosis transmembrane
conductance regulator (CFTR) gene on
chromosome 7. Most commonly, the
mutation in the CFTR gene is a three-
base-pair deletion. This protein is required
to regulate the components of sweat,
digestive fluids, and mucus. CFTR regulates
the movement of chloride and sodium ions
across epithelial membranes, such as the
alveolar epithelia located in the lungs.
Since all of the cells of a CF patient have
the defective protein, large quantities of
thick, sticky mucus build up throughout the
lungs and other organs. This results in the
severity of symptoms seen in CF patients.

Is It A Good Target For Gene Therapy?

To check this some questions must be answered:

Does the condition result from mutation? Yes.

Is the biology of the disorder known? Yes.

Will adding a normal copy of the gene fix the problem in the affected tissue? Yes.
While the mutated CFTR gene encodes a non-functional ion channel
protein, it will not prevent a normal CFTR channel protein from
working properly. Therefore, adding a normal copy of the CFTR gene
should fix the problem

Is it feasible to deliver the gene to the cells of the affected tissue? Yes, in part.
Treating the lungs of patients with CF might be feasible, since the
lung surfaces are exposed to the air and somewhat easy to reach.
Because the digestive system is less accessible, however, it might
be a more difficult region to treat.

Hence we can conclude that it is a perfect disease to be treated by gene


Choosing vectors
The vectors that are most suitable for gene therapy are:

Retroviruses are enveloped viruses that replicate in a host cell
through the process of reverse transcription. It is a single-stranded
RNA virus that stores its nucleic acid in the form of an mRNA
genome targets. Once inside the host cell cytoplasm the virus uses
its own reverse transcriptase enzyme to produce DNA from its RNA
genome, the reverse of the usual pattern, thus retro (backwards).
This new DNA is then incorporated into the host cell genome by an
integrase enzyme, at which point the retroviral DNA is referred to as
a provirus. The host cell then treats the viral DNA as part of its own
genome, translating and transcribing the viral genes along with the
cell's own genes, producing the proteins required to assemble new
copies of the virus.

One drawback of retroviruses, such as the Moloney retrovirus,
involves the requirement for cells to be actively dividing for
transduction. As a result, cells such as neurons are very resistant to
infection and transduction by retroviruses.

But the airway cells which are affected by the disease cystic
fibrosis and must be targeted divide infrequently. Hence Retrovirus
is not a suitable vector for this disease.

Adenoviruses (members of the family
Adenoviridae) are medium-sized (90
100 nm), nonenveloped (without an
outer lipid bilayer) viruses with
anicosahedral nucleocapsid
containing a double stranded DNA

They have a broad range of vertebrate

hosts and have been found to cause a wide range of illnesses, from
mild respiratory infections in young children to life-threatening
multi-organ disease in people with a weakened immune system.

But they can cause/induce an immune response in the human body

hence not suitable for gene delivery.

Herpes Simplex Virus

Herpes simplex viruses, also known

as Human herpes virus, are members
of the herpes virus family,
Herpesviridae, that infect humans.
They can be spread when an infected
person is producing and shedding the

virus. Herpes Simplex can be spread through contact with saliva,
such as sharing drinks.

But these viruses only affect the cells of the nervous system and
cannot infect the airway cells and hence not suitable.

Adeno-Associated Viruses

Adeno-associated virus (AAV) is a small virus which infects humans

and some other primate species. AAV is not currently known to
cause disease and consequently the virus causes a very
mild immune response.AAV can infect both dividing and quiescent
cells and persist in an extra chromosomal state without integrating
into the genome of the host cell. Despite its few disadvantages
these features make AAV a very attractive candidate for creating
viral vectors for gene therapy, and for the creation of isogenic
human disease models

Hence it is the best choice for gene delivery in the case of Cystic Fibrosis.

History of Cystic Fibrosis Gene Therapy

Gene therapy for cystic fibrosis began in 1990, when scientists
successfully corrected faulty cystic fibrosis transmembrane
conductance regulator (CFTR) genes. They did this by adding normal
copies of the gene to laboratory cell cultures.

In 1993, the first experimental CF gene therapy treatment was given
to a patient with cystic fibrosis. Researchers modified a common
cold Adenovirus to act as a delivery vehicle by carrying normal
genes to the CFTR cells in the nasal passages. Researchers chose
nasal passages as the site of delivery because they are easier to
access and measure gene activity than the lung airway. Later trials
delivered the vector to patients lung airways.

In the earlier trials, it had looked like the virus had entered cells and
that the CTFR gene was working. But later trials with different
patients showed levels of VFTR gene activity that were too low to
make any difference. Researchers came to think that the adenovirus
cant easily enter airway cells, especially in the low doses that were
being given. In the earlier trials, they speculated, gene activity
resulted from the damage to the cells during delivery allowing the
virus to enter easily.

Hence when higher doses of the virus were tried, the immune
system of the patients started mounting immune responses and
fighting off the virus. This caused a blockage in the trials until 1998.

Trials using Adeno-associated virus to deliver the CTFR gene began in
1998. Unlike the adenovirus, the Adeno-associated virus caused no
immune response or adverse side effects in patients.

But unlike the researchers predictions, the adeno-associated virus did not
enter cells efficiently and integrate into calls genomic DNA. They
produced only low and fleeting amounts of CFTR gene activity.
Researchers are still working to figure out what caused the viruses to fail.

But because it is safe, the virus as we predicted earlier holds promise

for being a good way to deliver the CFTR gene to patients airway cells.
But researchers need to learn more about how the virus infects cells in
order or make it an effective delivery method.

Some the factors that have kept gene therapy from becoming an
effective treatment for genetic diseases are:

Short-lived nature of gene therapy - Before gene therapy can

become a permanent cure for any condition, the therapeutic
DNA introduced into target cells must remain functional and
the cells containing the therapeutic DNA must be long-lived

and stable. Problems with integrating therapeutic DNA into the
genome and the rapidly dividing nature of many cells prevent
gene therapy from achieving any long-term benefits. Patients
will have to undergo multiple rounds of gene therapy.

Immune response -Anytime a foreign object is introduced into

human tissues, the immune system is designed to attack the
invader. The risk of stimulating the immune system in a way
that reduces gene therapy effectiveness is always a potential
risk. Furthermore, the immune system's enhanced response to
invaders it has seen before makes it difficult for gene therapy
to be repeated in patients.

Problems with viral vectors - Viruses, while the carrier of choice

in most gene therapy studies, present a variety of potential
problems to the patient --toxicity, immune and inflammatory
responses, and gene control and targeting issues. In addition,
there is always the fear that the viral vector, once inside the
patient, may recover its ability to cause disease.

Multigene disorders -Conditions or disorders that arise from

mutations in a single gene are the best candidates for gene
therapy. Unfortunately, some the most commonly occurring
disorders, such as heart disease, high blood pressure,
Alzheimer's disease, arthritis, and diabetes, are caused by the
combined effects of variations in many genes. Multigene or
multifactorial disorders such as these would be especially
difficult to treat effectively using gene therapy.

Issues regarding
Gene Therapy
What are the possible implications of gene therapy research to
society? All of us - researchers, policymakers and the public - have a

responsibility to explore the potential effects of gene therapy
research on our lives so that we can make informed decisions.

For each new application of gene therapy research, we must


What are the benefits?

What are the risks?

Whom will the technology help? Who will it potentially hurt?

What does gene therapy mean for us?

There are several types of issues to consider as we think about

gene therapy:

Ethical issues ask us to consider the potential moral outcomes

of gene therapy research.

Legal issues require researchers and the public to help

policymakers decide whether and how gene therapy research
should be regulated by the government.

Social issues involve the impact of gene therapy research on

society as a whole.

Some questions to ponder

When should gene therapy be used? Should it be used to treat

critically ill patients? Should it be used to treat babies and

What effect would gene therapy have on future generations if germline

(reproductive) cells were genetically altered? How might this alteration affect
human variation?

Who should decide what are "good" or "bad" uses of genetic

modifications? How do you define "normal" with regard to human

What if we could alter human traits not associated with disease? Would it be
okay to use gene therapy to improve or enhance a person's genetic

Who will have access to gene therapy, treatments and long-term follow-
ups? Will gene therapy and genetic enhancements create an
advantage for those who can afford it?

Recent Upcoming
CRISPR stands for clustered
regularly interspaced short
palindromic repeats. These RNA
sequences serve an immune
function in archaea and bacteria,
but in the last year or so,
scientists have seized upon them
to rewrite genes. The RNA
sequence serves as a guide to
target a DNA sequence in, say, a
zygote or a stem cell. The guide
sequence leads an enzyme,
Cas9, to the DNA of interest. Cas9 can cut the double strand, nick it,
or even knock down gene expression. After Cas9 injures the DNA,
repair systems fix the sequence - or new sequences can be

It isn't the first or only method of gene repair therapy thats been
developed, but the CRISPR technology, says Ramesar, is so special
because, unlike previous methods which were more laborious and
could only target one kind of cell in the body, it appears to be a "one
size fits all delivery", adaptable for different tissues. The procedure
also seems relatively simple to perform.
Exciting as the development may be, CRISPR wont be delivering
instant cures just yet.
Ramesar says, from his initial impressions of the literature, that it
would seem that localised, accessible abnormal tissue (as in the
retina or skin) could be targeted more easily.

Conditions affecting the body more systemically, however, such as
certain developmental syndromes, or central nervous system
disorders, might be problematic in terms of getting the repair
technology into enough of the target cells in that tissue to make an
effective difference.
"It may also depend on the stage one attempts to carry out the
therapy, in terms of the patients age and level of advancement of
the disease," says Ramesar.

Although early clinical failures led many to dismiss gene therapy as
over-hyped, clinical successes since 2006 have bolstered new
optimism in the promise of gene therapy. These include successful
treatment of patients with the retinal disease Leber's congenital
amaurosis, X-linked SCID, ADA-SCID, adrenoleukodystrophy, chronic
lymphocytic leukaemia (CLL),acute lymphocytic
leukaemia (ALL),multiple myeloma, haemophilia and Parkinson's
disease. These recent clinical successes have led to a renewed
interest in gene therapy, with several articles in scientific and
popular publications calling for continued investment in the field.




12th NCERT Biology
Stryer Biochemistry