遗传突变修复,基因治疗 z-bo 2020-08-17 542

　　The Juan Carlos Izpisua Belmonte laboratory of the Salk Institute in the United States, the BGI Li Yingrui team and the Liu Guanghui research group of the Institute of Biophysics of the Chinese Academy of Sciences collaborated for the first time to use whole-genome sequencing (WGS) to clarify the safety of existing disease genome targeted correction tools It is reliable and has created a new type of human genetic mutation repair tool telHDAdV that is far more efficient than the current genome-targeted editing technology, providing an important theoretical basis for the development of stem cell-based gene therapy.

　　The emergence of human induced pluripotent stem cell technology (iPSC) has promoted the rapid development of human disease genome targeted correction technology. Currently available methods for targeted correction of human disease genome include: ribozyme-mediated DNA homologous recombination technology (such as ZFN, TALEN, CRISPR/CAS9, etc.) and large-segment DNA homologous recombination technology that does not rely on ribozymes (with The third-generation adenovirus vector HDAdV is representative). The genetically repaired autologous stem cells have the potential to treat their own diseases, so they have broad application prospects in individual medicine and regenerative medicine.

　　Liu Guanghui’s research team was the first to use HDAdV-mediated genome-targeted editing technology to achieve targeted repair of the disease-causing gene LMNA in iPSCs of patients with progeria, conceptually confirming the feasibility of in situ correction of genetic mutations in patient cells . They then corrected the pathogenic mutations in the stem cells of patients with Parkinson's disease and Fanconi anemia, laying the foundation for the mechanism research, drug evaluation, and personalized stem cell and gene therapy of these genetic diseases.

　　In this study, the researchers used HDAdV, TALEN and CRISPR three different methods for the first time to target the mutated hemoglobin gene (HBB) in iPSC of patients with sickle cell anemia. It is found that these three gene correction methods have similar targeting efficiency for HBB gene. At the same time, the results of whole-genome deep sequencing show that TALEN and HDAdV maximize the integrity of the patient's genome during the gene correction process, which indicates the safety and reliability of these methods.