Figure 1 : General procedure of plant genome editing using CRISPR–Cas9.

Plant genome editing can usually be divided into 4 consecutive steps and shows the estimated time required for each step. PCR / RE, polymerase chain reaction / restriction enzyme digestion

Genetic editing includes CRISPR CAS 9: Simple version of the CRISPR / Cas system CRISPR / Cas 9 is a technique for comparatively easy genome editing. Scientists including co-inventors of CRISPR strongly called for global stops in the application of CRISPR to human germline, especially in clinical applications. "Scientists should avoid trying to use germline genomic alterations for human clinical application in a relaxed jurisdiction," he said. These scientists support the basic research of CRISPR and do not believe that the development of CRISPR is sufficient for all clinical use and can be genetically altered in humans.

Genomic editing with CRISPR / Cas9 has become an effective and reliable way to add accurate, targeted changes to the genome of living cells. CXCR4 is a co-receptor for human immunodeficiency virus type 1 (HIV - 1) infection and is thought to be an important therapeutic target for AIDS. CXCR4 mediates viral entry into human CD4 + cells by binding to the envelope protein gp120. Here we show that the human CXCR4 gene is efficiently disrupted by CRISPR / Cas9-mediated genomic editing, resulting in HIV-1 resistance in human primary CD4 + T cells. We also show that excision of Cas9-mediated CXCR4 shows high specificity and negligible off-target effect without affecting cell division and proliferation. Accurate and effective genome editing of CXCR4 provides a novel strategy for therapeutic application of HIV - 1 infection

The CRISPR / Cas 9 system has been used to create a simple RNA programmable method to mediate genomic editing in mammalian cells, and is used to generate gene knockouts (by insertion / deletion) or knock-in (by HDR) Can be used. In order to cause gene disruption (Figure 1), a single guide RNA (sgRNA) is generated to direct the Cas9 nuclease to a specific genomic location. Cas9-induced double-strand breaks are repaired by the NHEJ DNA repair pathway Repair is prone to mistakes and can cause insertions or deletions that may destroy gene function (INDEL)