A CRISPR-Cas9-triggered strand displacement amplification method for ultrasensitive DNA detection

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalNot applicablepeer-review

3 Scopus Citations
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Author(s)

  • Wenhua Zhou
  • Li Hu
  • Liming Ying
  • Zhen Zhao
  • Xue-Feng Yu

Detail(s)

Original languageEnglish
Article number5012 (2018)
Journal / PublicationNature Communications
Volume9
Early online date27 Nov 2018
Publication statusE-pub ahead of print - 27 Nov 2018

Link(s)

Abstract

Although polymerase chain reaction (PCR) is the most widely used method for DNA amplification, the requirement of thermocycling limits its non-laboratory applications. Isothermal DNA amplification techniques are hence valuable for on-site diagnostic applications in place of traditional PCR. Here we describe a true isothermal approach for amplifying and detecting double-stranded DNA based on a CRISPR-Cas9-triggered nicking endonuclease-mediated Strand Displacement Amplification method (namely CRISDA). CRISDA takes advantage of the high sensitivity/specificity and unique conformational rearrangements of CRISPR effectors in recognizing the target DNA. In combination with a peptide nucleic acid (PNA) invasion-mediated endpoint measurement, the method exhibits attomolar sensitivity and single-nucleotide specificity in detection of various DNA targets under a complex sample background. Additionally, by integrating the technique with a Cas9-mediated target enrichment approach, CRISDA exhibits sub-attomolar sensitivity. In summary, CRISDA is a powerful isothermal tool for ultrasensitive and specific detection of nucleic acids in point-of-care diagnostics and field analyses.

Research Area(s)

Citation Format(s)

A CRISPR-Cas9-triggered strand displacement amplification method for ultrasensitive DNA detection. / Zhou, Wenhua; Hu, Li; Ying, Liming; Zhao, Zhen; Chu, Paul K.; Yu, Xue-Feng.

In: Nature Communications, Vol. 9, 5012 (2018), 27.11.2018.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalNot applicablepeer-review

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