Cas-cleavable reporter systems,
CRISPR-Cas systems thereof, and methods of use thereof in
CRISPR-Cas based diagnostics. Cas-cleavable reporter systems may generate a luminescent, fluorescent, or other detectable
signal upon Cas-collateral cleavage of one or more reporter
system components.
CRISPR-Cas systems may comprise a Cas
protein having collateral cleavage activity, guide molecules, and the Cas-cleavable reporter
system. Cas-cleavable reporter systems may be a split
luciferase reporter
system, at least one element of which is bound to beads. For
multiplexing, CRISPR-Cas systems may comprise a Cas-cleavable quenched reporter system, optionally a Cas-cleavable quenched
fluorescent reporter system, a Cas
protein having collateral cleavage activity, and target-specific guide molecules attached to color-coded beads. CRISPR-Cas systems may further comprise amplification reagents. Methods may apply said CRISPR-Cas systems to
flow cell, well-plate, or other devices, and may measure detectable signals by microscopic,
plate reader, or other methods.