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Essential Features and Rational Design of CRISPR RNAs that Function with the Cas RAMP Module Complex to Cleave RNAs

机译:具有Cas RAMP模块复合体以分裂RNA的CRISPR RNA的基本特征和合理设计

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Small RNAs target invaders for silencing in the CRISPR-Cas pathways that protect bacteria and archaea from viruses and plasmids. The CRISPR RNAs (crRNAs) contain sequence elements acquired from invaders that guide CRISPR-associated (Cas) proteins back to the complementary invading DNA or RNA. Here, we have analyzed essential features of the crRNAs associated with the Cas RAMP module (Cmr) effector complex, which cleaves targeted RNAs. We show that Cmr crRNAs contain an 8 nucleotide 5' sequence tag (also found on crRNAs associated with other CRISPR-Cas pathways) that is critical for crRNA function and can be used to engineer crRNAs that direct cleavage of novel targets. We also present data that indicate that the Cmr complex cleaves an endogenous complementary RNA in Pyrococcus furiosus, providing direct invivo evidence of RNA targeting by the CRISPR-Cas system. Our findings indicate that the CRISPR RNA-Cmr protein pathway may be exploited to cleave RNAs of interest.
机译:小RNA靶向入侵者,使其在CRISPR-Cas途径中沉默,从而保护细菌和古细菌免受病毒和质粒的侵害。 CRISPR RNA(crRNA)包含从入侵者那里获得的序列元件,这些元件将CRISPR相关(Cas)蛋白引导回互补的入侵DNA或RNA。在这里,我们分析了与Cas RAMP模块(Cmr)效应子复合物相关的crRNA的基本特征,该复合物可裂解目标RNA。我们显示Cmr crRNA包含8个核苷酸的5'序列标签(也在与其他CRISPR-Cas途径相关的crRNA上发现),这对crRNA功能至关重要,可用于工程化直接切割新靶标的crRNA。我们还提供了表明Cmr复合物在激烈热球菌中切割内源性互补RNA的数据,提供了由CRISPR-Cas系统靶向RNA的直接体内证据。我们的发现表明,CRISPR RNA-Cmr蛋白途径可用于裂解目标RNA。

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