TALEN Targeted Gene Knockout Molecular Genetic Analysis

Introduction

Genome manipulation is a powerful technology, which can be used to specifically modify endogenous chromosomal DNA. This approach can be used to destroy or inactivate a gene, repair adverse mutations or increase the diversity of microbial population. With the help of this technology, various microorganisms can be genetically modified to create engineering microorganisms that meet our human needs and benefit human beings. It can also be used in fields such as synthetic biology, gene directed interference or multiple gene network interference, and gene therapy. Currently, we can provide microbial genome transformation, TALEN targeted gene knockout, Cas9 system gene knockout and other technical services.

The Process of the Gene Manipulation

1. To isolate DNA from the organism.
2. To put the DNA into the DNA vector.
3. To transfer the vector by transfection or transformation into the host.

•  Isolation of the DNA

We can get the part of the DNA thanks to restriction endonucleases - type II. The DNA strand is broken up in the specific places which are bordered by short nucleotide sequences. Both strands are cut out. Then we can recognize them and categorize by their size by agarose gel electrophoresis.

• Joining the DNA

The DNA ligase governs the properties of the segment. The cohesive ends are created and the insertion can takes place.

• Transfer into the Host

It is absolutelly necessary to choose the right vector for the DNA. It must accept the foreign DNA and continues its cell cycle. The most common are bacteria – especially E.coli. The transformation of DNA from a virus is called transfection. We have ensured that the integration into the host genome will be successful. The most important is to maintain the ability to replicate DNA.

The Complications of Use

The use of gene manipulation is difficult. It is a process that can change the human genes. So it brings many ethical problems. The safety is still discussable. The scientists are worried about the genetic diversity, because all of the modified organisms will be the same.