Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6
Author(s)Kellner, Max J.; Gootenberg, Jonathan S; Abudayyeh, Omar Osama; Joung, Julia; Collins, James J.; Zhang, Feng; ... Show more Show less
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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.
DepartmentInstitute for Medical Engineering and Science; Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences; Massachusetts Institute of Technology. Department of Mathematics; Massachusetts Institute of Technology. Department of Mechanical Engineering
American Association for the Advancement of Science (AAAS)
Gootenberg, Jonathan S., Omar O. Abudayyeh, Max J. Kellner, Julia Joung, James J. Collins, and Feng Zhang. “Multiplexed and Portable Nucleic Acid Detection Platform with Cas13, Cas12a, and Csm6.” Science 360, no. 6387 (February 15, 2018): 439–444.
Author's final manuscript