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CDetection: CRISPR-Cas12b-based DNA detection with sub-attomolar sensitivity and single-base specificity

机译:CDetection:基于CRISPR-Cas12b的DNA检测具有亚等摩尔灵敏度和单碱基特异性

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摘要

CRISPR-based nucleic acid detection methods are reported to facilitate rapid and sensitive DNA detection. However, precise DNA detection at the single-base resolution and its wide applications including high-fidelity SNP genotyping remain to be explored. Here we develop a Cas12b-mediated DNA detection (CDetection) strategy, which shows higher sensitivity on examined targets compared with the previously reported Cas12a-based detection platform. Moreover, we show that CDetection can distinguish differences at the single-base level upon combining the optimized tuned guide RNA (tgRNA). Therefore, our findings highlight the high sensitivity and accuracy of CDetection, which provides an efficient and highly practical platform for DNA detection.Electronic supplementary materialThe online version of this article (10.1186/s13059-019-1742-z) contains supplementary material, which is available to authorized users.
机译:据报道,基于CRISPR的核酸检测方法可促进快速,灵敏的DNA检测。然而,以单碱基分辨率进行精确的DNA检测及其包括高保真SNP基因分型在内的广泛应用仍有待探索。在这里,我们开发了一种Cas12b介导的DNA检测(CDetection)策略,与先前报道的基于Cas12a的检测平台相比,该策略对被检测的靶标显示出更高的灵敏度。此外,我们表明CDetection通过结合优化的优化的指导RNA(tgRNA)可以区分单碱基水平的差异。因此,我们的发现突出了CDetection的高灵敏度和准确性,这为DNA检测提供了一个高效且高度实用的平台。电子补充材料本文的在线版本(10.1186 / s13059-019-1742-z)包含补充材料,其中可供授权用户使用。

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