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.
Source: Nature Communications
Published by GSM · USA (original Chinese edition)
Photo: 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.
