艾滋病,基因编辑技术 z-bo 2020-10-09 345

　　The CRISPR/Cas9 system provides a new potential tool for editing the HIV-1 virus genome. Recently, researchers from the Infectious Disease Center of the Kobe University School of Medicine and the International School of Health of the Institute of Health Sciences designed RNA guidelines. CRISPR/Cas9 targets HIV-1 regulatory genes tat and rev. All guide RNAs (gRNAs) are based on the specific design of CRISPR, and the target sequence is conserved among the six major HIV-1 subtypes. Before co-transfection, each gRNA was cloned into CRISPRv2 lentivirus to create a lentiviral vector and transfect it into the cell. Is CRISPR/Cas9 293 compared to cells transfected or untransfected with an empty vector? It stably expresses Tat and Rev in T and HeLa cells, and usually inhibits the expression of these two genes in cells.

　　tat function test shows that transfection with tat-CRISPR can significantly inhibit the expression of luciferase driven by the HIV-1 promoter, while the Rev function test shows that gp120 transfected by rev-CRISPR. The results showed that the expression was completely suppressed. The target genes at the Cas9 splice site have high frequency mutations of varying degrees. It is worth noting that the researchers did not detect mutations at non-target sites, and Cas9 expression did not affect cell viability.

　　The researchers further tested the CRISPR/Cas9 system in HIV-1 infected T cell lines, and the re-stimulated cytokines significantly inhibited p24 expression, and used 6 gRNAs at the same time. Then I found that it can be further enhanced. Therefore, using the CRISPR/Cas9 system to target HIV-1 regulatory genes may be an effective way to achieve functional cure.