拆分向导 RNA 让 Cas9 获得核酸检测能力 30分钟内皮摩尔级灵敏度、单核苷酸特异性
《自然·通讯》11月14日发表一项 CRISPR 检测研究。研究团队把单向导 RNA 拆分为功能独立的支架模块和间隔模块，构建出拆分 RNA 向导 Cas9 系统，简称 SCas9。
CRISPR-Cas9 本身的反式切割活性有限，长期制约其在核酸检测中的应用。SCas9 克服了这一限制，可在无需扩增的情况下检测单链 DNA 和 RNA，灵敏度达到皮摩尔级，30分钟内完成，并具有单核苷酸特异性。
研究还引入 λ 外切酶，使系统能够对双链 DNA 进行不依赖 PAM 序列的检测，并在临床样本检测中达到阿摩尔级灵敏度。
论文题为《用于不依赖 PAM 且无顺式切割的核酸检测的拆分 RNA 向导 Cas9 系统》，由 Yin、Xu、Xia 等作者共同发表。论文信息同时披露，湖北大学与 BravoVax 有两项与该工作相关的 CRISPR/Cas9 检测技术专利申请在审，作者 YL 与 JQ 为发明人。
Split RNA-Guided Cas9 System SCas9 Enables Nucleic Acid Detection Within 30 Minutes at Picomolar Sensitivity
Nature Communications on November 14 published a CRISPR diagnostics study presenting a split RNA-guided Cas9 system, SCas9, created by partitioning the single-guide RNA into functionally independent scaffold and spacer modules.
The CRISPR-Cas9 system exhibits limited trans-cleavage activity, restricting its utility for nucleic acid detection. SCas9 overcomes this limitation, enabling amplification-free detection of single-stranded DNA and RNA with picomolar sensitivity within 30 minutes and single-nucleotide specificity.
Incorporation of λ exonuclease permits PAM-independent detection of double-stranded DNA, and the system achieved attomolar sensitivity in clinical assays described in the paper.
The paper, 'A split RNA-guided Cas9 system for PAM-independent and cis-cleavage-free nucleic acid detection', is authored by Yin, Xu, Xia and colleagues. The article also discloses that the regents of Hubei University and BravoVax have two patent applications relating to the work pending for CRISPR/Cas9 detection technologies, with authors YL and JQ as inventors.
来源：《自然·通讯》
发布：GSM·美国（发布链接）
图片来源：图：Cas9 蛋白与向导 RNA、靶 DNA 复合物的结构渲染图（资料图）。来源：Synpath，Wikimedia Commons，CC0。 / Photo: Structural rendering of Cas9 bound to guide RNA and target DNA (file image). Source: Synpath, Wikimedia Commons, CC0.
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