New CRISPR tool has been released by Chinese scientists

Recently, the international academic journal Nature Methods published the latest research results of the Targeted AID-mediated mutagenesis (TAM) enablesefficient genomic diversification in mammalian cells by the Changxing Research Group of the Shanghai Institutes for Biological Sciences / Shanghai Jiaotong University School of Medicine and Institute of Health Sciences. They combined activation-induced cytidine deaminase (AID) with CRISPR-dCas9 and used recombinant mouse proteins to create an effective genetic diversity tool for high-throughput screening of functional variants. This is a new method of DNA Base editing which is efficient and high-throughput.

Inspired by the "mutant self" mechanism and under the guidance of researcher Chang Xing, Dr. Ma Yunqing and Zhang Jiayuan discovered that when the nucleosidase-deficient Cas9 protein and the cytosine deaminase AID that induced antibody high-frequency mutation were fused, cytosine and guanine can be random to the other three bases on sgRNA-targeted genomic DNA. This new method can be specific to the cell DNA sequence diversity, the completion of genetic screening so as to analyze single nucleotide mutations function. At the same time, under the assist of a peptide inhibitor, dCas9-AID can induce specific cytosine to thymidine conversion to achieve precise editing of single base. The researchers used dCas9-AIDx to target BCR-ABL in chronic myelogenous leukemia cells, which effectively identified known mutations and new mutations that confer resistance to imatinib. Studies have shown that dCas9-AID is a very useful genetics tool and can screen functional acquired variation at the single-base level.

The team further demonstrated that this approach could rapidly and effectively mimics the heterogeneity of drug resistance in tumor cells, predict possible mutations in drug resistance, thus improving small molecule inhibitors and studying interaction of small molecules and protein targets. The results of the study provide a new approach of analysis of gene regulatory elements at the level of molecular evolution, gene therapy and single base level. Flarebio offers recombinant proteins of good quality such as recombinant CDH12 at good prices.