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Rapid characterization of anti-CRISPR proteins and optogenetically engineered variants using a versatile plasmid interference system

Nucleic Acids Research. 2023-10; 
Guoxu Song, Chunhong Tian, Jiahui Li, Fei Zhang, Yuxin Peng, Xing Gao and Yong Tian
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Gene Synthesis DNA sequences encoding Acr proteins were synthesized by Biomed or Genscript and ligated into pBAD24 vectors. Get A Quote


Anti-CRISPR (Acr) proteins are encoded by mobile genetic elements to overcome the CRISPR immunity of prokaryotes, displaying promises as controllable tools for modulating CRISPR-based applications. However, characterizing novel anti-CRISPR proteins and exploiting Acr-related technologies is a rather long and tedious process. Here, we established a versatile plasmid nterference with CRISPR interference (PICI) system in Escherichia coli for rapidly characterizing Acrs and developing Acr-based technologies. Utilizing the PICI system, we discovered two novel type II-A Acrs (AcrIIA33 and AcrIIA34), which can inhibit the activity of SpyCas9 by affecting DNA recognition of Cas9. We further constructed a circularly per... More