Zhao Li, You Di, Wang Ting, Zou Zhen-Ping, Yin Bin-Cheng, Zhou Ying, Ye Bang-Ce
Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Institute of Engineering Biology and Health, Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences, Zhejiang University of Technology, Hangzhou 310014, Zhejiang, China.
PNAS Nexus. 2022 Dec 6;1(5):pgac277. doi: 10.1093/pnasnexus/pgac277. eCollection 2022 Nov.
CRISPR-Cas, the immune system of bacteria and archaea, has been widely harnessed for genome editing, including gene knockouts and knockins, single-base editing, gene activation, and silencing. However, the molecular mechanisms underlying fluctuations in the genome editing efficiency of crispr in various cells under different conditions remain poorly understood. In this work, we found that Cas9 can be ac(et)ylated by acetyl-phosphate or acyl-CoA metabolites both and . Several modifications are associated with the DNA or sgRNA binding sites. Notably, ac(et)ylation of Cas9 driven by these metabolites in host cells potently inhibited its binding and cleavage activity with the target DNA, thereby decreasing Crispr genome editing efficiency. This study provides more insights into understanding the effect of the intracellular environment on genome editing application of crispr with varying efficiency in hosts.
CRISPR-Cas是细菌和古细菌的免疫系统，已被广泛用于基因组编辑，包括基因敲除和敲入、单碱基编辑、基因激活和沉默。然而，在不同条件下各种细胞中CRISPR基因组编辑效率波动背后的分子机制仍知之甚少。在这项工作中，我们发现Cas9可以被乙酰磷酸或酰基辅酶A代谢物乙酰化，且这两种代谢物都与DNA或sgRNA结合位点相关。值得注意的是，宿主细胞中由这些代谢物驱动的Cas9乙酰化会有效抑制其与靶DNA的结合和切割活性，从而降低CRISPR基因组编辑效率。这项研究为理解细胞内环境对宿主中效率各异的CRISPR基因组编辑应用的影响提供了更多见解。
