Abstract

This work describes a modification of the pRH2521 vector of the pRH2502/pRH2521 system for CRISPR-dCas9-mediated RNA interference. The modification enabled an increase in the cloning efficiency of guide RNA spacers. The ability of the modified pRH2502/pRH2521 system to suppress the transcription of certain genes was evaluated with the use of genes of Mycobacterium tuberculosis adenylate cyclases. The results revealed the limitations of the pRH2502/pRH2521 system for CRISPR interference associated with the probability of the detection of a protospacer adjacent motif (PAM) in the gene promoter region.

中文翻译：

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来自结核分枝杆菌的腺苷酸环化酶的 CRISPR 干扰

摘要

这项工作描述了对 pRH2502/pRH2521 系统的 pRH2521 载体的修改，用于 CRISPR-dCas9 介导的 RNA 干扰。该修改提高了向导 RNA 间隔区的克隆效率。使用结核分枝杆菌腺苷酸环化酶的基因评估了修饰的 pRH2502/pRH2521 系统抑制某些基因转录的能力。结果揭示了 pRH2502/pRH2521 系统对 CRISPR 干扰的局限性，与在基因启动子区域检测到原型间隔区相邻基序 (PAM) 的概率相关。