GENE THERAPY

Gene therapy is a method for treating genetic diseases. It is based on the principle that faulty genes, those with mutated
DNA sequences, can be replaced with the "correct" gene within a cell. Therefore, the disease can be stopped or cured.
In the future, this technique may allow doctors to treat a disorder by inserting a gene into a patients cells instead of
using drugs or surgery. Researchers are testing several approaches to gene therapy, including:

Replacing a mutated gene that causes disease with a healthy copy of the gene.

Inactivating, or knocking out, a mutated gene that is functioning improperly.

Introducing a new gene into the body to help fight a disease.

Although gene therapy is a promising treatment option for a number of diseases (including inherited disorders, some types
of cancer, and certain viral infections), the technique remains risky and is still under study to make sure that it will be safe
and effective. Gene therapy is currently only being tested for the treatment of diseases that have no other cures.
Types of Gene Therapy
1. Somatic Cell Gene Therapy
Therapeutic genes transferred into the somatic cells
Eg. Introduction of genes into bone marrow cells, blood cells, skin cells etc.
Will not be inherited by later generations
2. Germ Line Gene Therapy
Therapeutic genes transferred into the germ cells.
Eg. Genes introduced into eggs and sperms

Inherited and passed on to later generations.

3. 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.
4. 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.

Modes of Gene Therapy EX VIVO GENE THERAPY

1. In vivo gene therapy
direct delivery of genes into the cells of
a particular tissue in the body
2. Ex vivo gene therapy
transfer of genes to cultured cells and
reinsertion.
 FIGURE 1. Using gene therapy to cure a retinal degenerative disease in dogs
Researchers were able to use genes from healthy dogs to restore vision in dogs blinded by an inherited retinal
degenerative disease. This disease also occurs in human infants and is caused by a defective gene that leads to early vision
loss, degeneration of the retinas, and blindness. In the gene therapy experiments, genes from dogs without the disease were
inserted into 3-month-old dogs that were known to carry the defective gene and that had been blind since birth. Six weeks
after the treatment, the dogs' eyes were producing the normal form of the gene's protein product, and by three months, tests
showed that the dogs' vision was restored.

BIOLISTIC TECHNOLOGY
Biolistic technology, also called particle bombardment, is a direct physical method of introducing nucleic acids into
cells. Nucleic acids or other biological molecules are coated onto high-density gold or tungsten micro particles (micro
carriers), which are then accelerated to high velocity by a helium pulse and driven through cell walls and membranes into
the target. The physical nature of this technology makes it extremely versatile and easy to use. It can be applied to wide
range of targets, including cell cultures, tissues, organs, plants, animals, and bacteria, as well as organelles.
A gene gun, or a biolistic particle delivery system, is a genetic engineering technique originally designed to modify
plants. It is used for delivery of exogenous DNA to cells. This method is known as 'biolistics'. Gene guns can be used
effectively on most cells but are mainly used on plant cells.
Step 1: The gene gun apparatus is ready to fire.
Step 2: Helium fills the chamber and pressure builds
against the rupture disk.
Step 3 :The pressure eventually reaches the point
where the rupture disk brakes, and the resulting burst
of helium propels the DNA/gold-coated macrocarrier
('Plastic Disk') into the stopping screen.
Step 4: When the macrocarrier hits the stopping
screen, the DNA-coated gold particles are propelled
through the screen and into the target cells.

Gene Gun

It was invented by three scientists from Cornell, John C Stanford, Ed Wolf,

and Nelson Allen, and one scientist from DuPont, Ted Klein, between the years
1983 and 1986.
This device injects cells with genetic information using a heavy metal
element bullet coated with plasmid DNA.
This device can be used on almost any cell, and it is not limited to
transforming genetic information in the nucleus. It also can modify other
organelles.
Originally, the gene gun was a Crosman air pistol that had been redesigned
to fire dense tungsten particles.
The gene gun today is used not only on plants cells, but also on bacteria,
yeast, and mammalian cell lines that had previously been impossible to inject
such as non-dividing cells or primary cells.
Using the gene gun on other organelles, such as chloroplast, has been
useful because no bacteria or viruses are known to infect the chloroplast.
Therefore, other ways of transfer genetic
DNA delivery with the gene gun offers new research opportunities for
areas such as DNA vaccination/genetic immunization, gene therapy, tumor
biology, and many others.