CRISPR-Cas9,基因编辑 z-bo 2020-09-24 554

　　Cellesearch is a subsidiary of Nature Publishing Group, a research team of Wang Haoyi from the Institute of Zoology, Chinese Academy of Sciences and Zhang Yongping from Beijing Genbu Hospital. We have published a research paper called "Online". Liu Xiaojuan, Zhang Yongping and Cheng Chen are the co-lead authors of the paper, and researcher Wang Haoyi is the author of the paper. CART (Chimeric Antigen Receptor T Cell) cell therapy is a very promising tumor treatment method, which is translated as "Chimeric Antigen Receptor T Cell" in Chinese.

　　Chimera Antigeneceptor (Chimera Antigeneceptor, CAR) is an artificially designed and synthesized transmembrane protein, which contains three parts: extracellular, intracellular membrane and intracellular. The extracellular part of CAR is mainly monoclonal antibody variable region (scFV), which can recognize specific antigens and is responsible for recognizing specific antigens. The transmembrane region ensures that the CAR is located on the cell membrane, while the intracellular region is involved in T cell activation and T cell-mediated immune response. Clinical data shows that CART has achieved excellent efficacy in the treatment of B-cell malignancies. However, all currently used methods are autologous cell transplantation (ACT), which is expensive and time-consuming. For newborns and elderly patients, it is difficult to obtain sufficient and high-quality lymphocytes for CAR T cell therapy. Therefore, the important research direction of CAR T cell therapy is to use T cells from healthy blood donors to prepare a large number of CAR T cells for clinical use by hundreds of patients. The establishment of this technology will greatly reduce the cost of trolley treatment, better guarantee the quality of prepared cells, and allow patients to immediately obtain trolley cells for treatment when needed.

　　Allogeneic donor T cells are used to prepare universal CART to treat multiple patients, so as to ensure its safety and prevent adoptive cells from attacking the patient's own cells, while immunogenicity should be reduced to avoid attack. Wang Haoyi's research team from the Institute of Animal Science of the Chinese Academy of Sciences used CRISPR-Cas9 technology to knock out the αβ T cell receptor (TCR) from allogeneic CAR T cells to avoid implantation host disease (GVHD). done. The knockout of human leukocyte antigen (HLA) reduces its own immunogenicity. In addition, PD-1 is a T cell surface inhibitor related to tumor immune escape. CART believes that blocking the PD-1 signaling pathway has produced good effects in many types of tumors. I use cells to knock out PD. -1 gene blocks the PD-1 signaling pathway. The preparation of allogeneic CART cells requires simultaneous editing of multiple genes, and the RNA-mediated DNA nuclease system CRISPR-Cas9 is a very suitable technology platform. In this study, two genes (TRAC and B2M) or three genes (TRAC, B2M and PD-1) in CAR T cells were knocked out using the CRISPR-Cas9 system.

　　"The results show that, compared with conventional CAR T cells, these gene-edited CAR T cells have the same or stronger tumor cell killing function in vitro and in vivo, making them effective effect cells for clinical applications. It shows the expected result. The combination of gene editing technology and adoptive immunity has a wide range of potential applications in the treatment of tumors and HIV/AIDS, and the establishment of this method has laid a solid technical foundation for the study of these diseases.