Principle of CRISPR-Cas9 technology: CRISPR-Cas9 technology contains two important components, one is Cas9 protein which performs the function of DNA double-stranded cleavage, and the other is gRNA which has the function of guiding. CRISPR-Cas9 technology uses gRNA complementary to the target sequence to guide Cas9 nuclease to recognize and cut the specifically targeted DNA, resulting in double-stranded or single-stranded DNA breakage. Then, cells will use their own two DNA repair mechanisms to repair the broken DNA, that is, non-homologous terminal splicing or homologous-mediated repair.
Use of CRISPR-Cas9: Gene insertion and knockout can be achieved by using two repair mechanisms. In the absence of template, using NHEJ repair, random insertion or deletion of sequences leads to gene knockout. When there is a template, the target modified gene can be introduced into the cleavage site through HDR repair to realize the fixed-point modification of the gene.
strong function
CRISPR-Cas9 is an adaptive immune defense formed by bacteria and archaea in the long-term evolution process, which can be used to fight against invading viruses and foreign DNA. CRISPR-Cas9 gene editing technology is a specific DNA modification technology for the target gene, which is also a cutting-edge method used in gene editing.
Gene editing technology based on CRISPR-Cas9 shows great application prospect in a series of application fields of gene therapy, such as hematological diseases, tumors and other genetic diseases. This technical achievement has been applied to the precise genome transformation of human cells, zebrafish, mice and bacteria.
Patent authorization
On April 5, 20 14, 14, the first patent authorization of CRISPR was obtained from the US Patent and Trademark Office. Patents include the use of CRISPR in eukaryotic cells or any nucleated species. This means the right to use CRISPR on all living things except bacteria, including mice, pigs and people.