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An inducible CRISPRi circuit for tunable dynamic regulation of gene expression in Saccharopolyspora erythraea

Biotechnol Lett. 2024-01; 
Xing Jiang, Xiang Ke, Xiwei Tian, Ju Chu
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Gene Synthesis … CymRP21 promoter was obtained from pUC57-CymRP21 by XbaI and NdeI digestion, which were synthesized according to the sequence provided by Horbal (Genscript.CN) (Horbal et … Get A Quote

摘要

Actinomyces are gram-positive bacteria known for their valuable secondary metabolites. Redirecting metabolic flux towards desired products in actinomycetes requires precise and dynamic regulation of gene expression. In this study, we integrated the CRISPR interference (CRISPRi) system with a cumate-inducible promoter to develop an inducible gene downregulation method in Saccharopolyspora erythraea, a prominent erythromycin-producing actinobacterium. The functionality of the cumate-inducible promoter was validated using the gusA gene as a reporter, and the successful inducible expression of the dCas9 gene was confirmed. The developed inducible CRISPRi strategy was then employed to downregulate the expression of ... More

关键词

Saccharopolyspora erythraea, CRISPRi, Dynamic regulation, Inducible promoter
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