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Current Protocols Essential Laboratory Techniques Pub Date : 2020-12-16 DOI:10.1002/cpet.34

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Abstract

Cover: In Sirois et al. (http://doi.org/10.1002/cpet.45), the image shows the CRISPR/Cas system. Bacteria and archaea use the CRISPR/Cas system for adaptive immunity against foreign viruses and plasmids. (1) Upon infection, foreign DNA is processed into small (∼20-bp) fragments which are (2) incorporated between palindromic repeats within the CRISPR locus. (3) The CRISPR locus, containing the spacers and trans-activating RNA (tracrRNA), is transcribed. (4) The spacer transcripts are processed to generate small CRISPR RNA (crRNA) molecules, which base pair with the tracrRNA to form a functional guide RNA (gRNA). (5) A Cas endonuclease protein complexes with the gRNA via the tracrRNA, and is guided to a specific target DNA by the crRNA. (6) Upon encountering the specific DNA sequence, the Cas protein will hybridize to, and cut, the double-stranded DNA, resulting in a double-strand break. Figure adapted from “CRISPR-Cas9 Adaptive Immune System of Streptococcus pyogenes against bacteriophages,” by BioRender.com (2020). Retrieved from https://app.biorender.com/biorender-templates.

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封面:在Sirois et al. (http://doi.org/10.1002/cpet.45)中，图像显示了CRISPR/Cas系统。细菌和古细菌利用CRISPR/Cas系统对外来病毒和质粒产生适应性免疫。(1)感染后，外源DNA被加工成小的(~ 20bp)片段，(2)在CRISPR位点内的回文重复序列之间合并。(3)包含间隔子和反式激活RNA (tracrRNA)的CRISPR位点被转录。(4)间隔转录本经过加工生成小分子CRISPR RNA (crRNA)，与tracrRNA碱基配对形成功能引导RNA (gRNA)。(5)一个Cas内切酶蛋白通过tracrRNA与gRNA复合物，并被crRNA引导到特定的靶DNA。(6)当遇到特定的DNA序列时，Cas蛋白会与双链DNA杂交并切割，导致双链断裂。图改编自BioRender.com(2020)的“抗噬菌体的化脓性链球菌的CRISPR-Cas9适应性免疫系统”。检索自https://app.biorender.com/biorender-templates。

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Current Protocols Essential Laboratory Techniques

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