The group of Chunbo LOU, together with the Peking iGEM team, developed a novel paired dCas9 reporter system. As a proof of concept, they successfully distinguished Mtb genome markers. The paired dCas9 reporter system is programmed by single guide RNAs complementary to the upstream and downstream segments of a target DNA sequence, respectively. When substrate DNA containing the two segment-sequences in proximity are detected and bound by a pair of dCas9, luminescence is generated from the catalytic activity of luciferase in its entirety. In this detection system, sequence information (including complementary sequence and spacer sequence) and output signal amplification are obtained simultaneously.
CAS news release, November 10, 2016