Article 1: Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6 Jonathan S. Gootenberg1,2,3,4,7,, Omar O. Abudayyeh1,2,3,4,5,

Article 1: Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6 Jonathan S. Gootenberg1,2,3,4,7,, Omar O. Abudayyeh1,2,3,4,5,, Max J. Kellner1, Julia Joung1,2,3,4, James J. Collins1,4,5,6,8, Feng Zhang1,2,3,4,† Abstract 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 pre-amplification 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 SHERLOCKv2: 1) 4-channel single reaction multiplexing using orthogonal CRISPR enzymes; 2) quantitative measurement of input down to 2 aM; 3) 3.5-fold increase in signal sensitivity by combining Cas13 with Csm6, an auxilary CRISPR-associated enzyme; and 4) lateral flow read-out. SHERLOCKv2 can detect Dengue or Zika virus ssRNA 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. http://science.sciencemag.org/content/early/2018/02/14/science.aaq0179 http://science.sciencemag.org/content/early/2018/02/14/scien...