Details

Autor(en) / Beteiligte

• Wang, Ting
• Liu, Yong
• Sun, Huan‐Huan
• Yin, Bin‐Cheng
• Ye, Bang‐Ce

Titel

An RNA‐Guided Cas9 Nickase‐Based Method for Universal Isothermal DNA Amplification

Ist Teil von

• Angewandte Chemie International Edition, 2019-04, Vol.58 (16), p.5382-5386

Auflage

International ed. in English

Ort / Verlag

Germany: Wiley Subscription Services, Inc

Erscheinungsjahr

2019

Quelle

MEDLINE

Beschreibungen/Notizen

• We have developed an ingenious method, termed Cas9 nickase‐based amplification reaction (Cas9nAR), to amplify a target fragment from genomic DNA at a constant temperature of 37 °C. Cas9nAR employs a sgRNA:Cas9n complex with a single‐strand nicking property, a strand‐displacing DNA polymerase, and two primers bearing the cleavage sequence of Cas9n, to promote cycles of DNA replication through priming, extension, nicking, and displacement reaction steps. Cas9nAR exhibits a zeptomolar limit of detection (2 copies in 20 μL of reaction system) within 60 min and a single‐base discrimination capability. More importantly, the underlying principle of Cas9nAR offers simplicity in primer design and universality in application. Considering the superior sensitivity and specificity, as well as the simple‐to‐implement, rapid, and isothermal features, Cas9nAR holds great potential to become a routine assay for the quantitative detection of nucleic acids in basic and applied studies. In good nick: A method, termed Cas9 nickase‐based amplification reaction (Cas9nAR), for the isothermal amplification of a target fragment from genomic DNA was developed. The method has a zeptomolar limit of detection and a single‐nucleotide discrimination capability.