INTRO

Hello everyone. Today I’m going to talk to you about the CRISPR CAS9
system. It's a new simple, fast and efficient system for cutting DNA at a
specific location in the genome, in any cell. It consists of a "guide RNA",
which targets a particular DNA sequence, associated with the enzyme
Cas9, which, like molecular scissors, cuts the DNA.

Simple to implement, this technique can be easily used by researchers

to better understand the role of genes, correct defective DNA, develop
new animal models, implement new therapeutic strategies, etc.

Once the DNA sequence is cut, the cell's repair systems will reattach the
ends of the two pieces of DNA created by the cut.
There are then 2 possibilities:

In the absence of a template sequence, the repair process adds or

removes a few nucleotides from each of the DNA ends in order to be
able to reattach them; this causes “abnormalities” in the targeted DNA
sequence: the gene then becomes randomly inactive or repaired;
In the presence of a synthetic DNA sequence without genetic anomaly
introduced by the researchers into the cell, the repair process integrates
it at the level of the cut: the gene is then repaired or corrected.

PART 1:A defense system against

viruses

The CRISPR/Cas9 system was discovered in bacteria. A large number

of them have in their genome short DNA sequences repeated regularly
along their genome, called CRISPR (for Clustered Regularly Interspaced
Short Palindromic Repeats). They allow bacteria to retain the memory of
an infection by a virus to better defend themselves against it the
following times.
Following infection by a virus, DNA sequences from the virus are
integrated into these CRISPR sequences. During a new infection by this
virus, the viral DNA within CRISPR is copied into RNA and is associated
with the Cas9 protein (for CRISPR associated). This RNA associated
with Cas9 binds to the DNA of the virus and Cas9 inactivates the viral
DNA by cutting it. The virus is thus eliminated.

PART 2:
It was in 2012 that the two researchers Emmanuelle Charpentier and
Jennifer Doudna succeeded in hijacking this biological system to turn it
into veritable molecular scissors capable of targeting specific locations in
the genome of any cell.
This technique is now used in thousands of research laboratories around
the world. The simplicity of its implementation allowed a very rapid
diffusion of its use within the scientific community.

Indeed, all that is needed is a Cas9 protein to cut the DNA and a guide
RNA specific to the targeted sequence. The guide RNA, which must be
adapted each time, is very easy to manufacture, especially since
software (free access) has been developed to determine the best
sequences to use depending on the gene or the sequence. targeted.

In October 2020, the two researchers Emmanuelle Charpentier and

Jennifer Doudna were awarded the Nobel Prize in Chemistry for the
development of these "molecular scissors".

PART 3:

However, much work is still needed to ensure the safety of use of this
approach in humans. In addition, the administration of the CRISPR/Cas9
system in the cells of the body will have to be carried out using vectors
and will come up against certain obstacles already encountered in gene
therapy: the choice of the vector (associated adenovirus, lentivirus ,
synthetic vector, etc.) or the control of the immune response.
As with all techniques for modifying the genome of living beings, the
ethical limits of the application of this approach also need to be defined.

Conclusion:

Although CRISPR-Cas9 has some flaws, its affordability and ease of use
make it a popular tool in research. Based on the recognition of a target
DNA from a guide RNA then by the double-strand cut of this DNA by
Cas9, it is mainly involved in genome editing. This genome editing is
applied in many fields, ranging from the food industry to the discovery of
new drugs. The application of these methods in humans has raised
many ethical questions, particularly regarding their use on embryos.
Indeed, this would totally change the genome of an individual, which
raises fears of potential abuses of eugenics.