Maqola

Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6

Jonathan S. GootenbergBroad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USAOmar O. AbudayyehBroad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USAMax J. KellnerBroad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USAJulia JoungBroad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USAJames J. CollinsBroad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USAFeng ZhangBroad Institute of the Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA

2018en

ABI

Annotatsiya

Rapid detection of nucleic acids is integral for clinical diagnostics and biotechnological applications. We recently developed a platform termed SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) that combines isothermal preamplification with Cas13 to detect single molecules of RNA or DNA. Through characterization of CRISPR enzymology and application development, we report here four advances integrated into SHERLOCK version 2 (SHERLOCKv2) (i) four-channel single-reaction multiplexing with orthogonal CRISPR enzymes; (ii) quantitative measurement of input as low as 2 attomolar; (iii) 3.5-fold increase in signal sensitivity by combining Cas13 with Csm6, an auxiliary CRISPR-associated enzyme; and (iv) lateral-flow readout. SHERLOCKv2 can detect Dengue or Zika virus single-stranded RNA as well as mutations in patient liquid biopsy samples via lateral flow, highlighting its potential as a multiplexable, portable, rapid, and quantitative detection platform of nucleic acids.

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Identifikatorlar

DOI: 10.1126/science.aaq0179

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