Acylation driven by intracellular metabolites in host cells inhibits Cas9 activity used for genome editing

Li Zhao; Di You; Ting Wang; Zhen-Ping Zou; Bin-Cheng Yin; Ying Zhou; Bang-Ce Ye

doi:10.1093/pnasnexus/pgac277

Acylation driven by intracellular metabolites in host cells inhibits Cas9 activity used for genome editing

PNAS Nexus. 2022 Dec 6;1(5):pgac277. doi: 10.1093/pnasnexus/pgac277. eCollection 2022 Nov.

Authors

Li Zhao¹, Di You¹, Ting Wang¹, Zhen-Ping Zou¹, Bin-Cheng Yin^{1

2}, Ying Zhou¹, Bang-Ce Ye^{1

2}

Affiliations

¹ Laboratory of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
² 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.

Abstract

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 in vitro and in vivo. 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.

Keywords: Cas9; Crispr; acylation; genome editing.