TEMPORAL CONTROL OF CRISPR-Cas9 ACTIVITY USING BIOORTHOGONAL CHEMISTRY

Abstract

Disclosed are systems and methods for reversible, chemically inducible regulation of CRISPR-Cas9 activity using bioorthogonal chemistry. The invention includes modified guide RNAs (sgRNAs) incorporating a first bioorthogonal reactive group that does not interfere with native Cas9 function. Upon introduction of a CRISPR suppressor comprising a second complementary bioorthogonal reactive group, a rapid and selective chemical reaction occurs under physiological conditions, resulting in inhibition of Cas9-mediated genome editing. The suppressors may include small molecules, peptides, or nucleic acid analogs designed to sterically or conformationally disrupt the ribonucleoprotein complex. This platform enables precise spatial and temporal control over CRISPR-Cas9 activity, with potential applications in improving editing specificity and reducing off-target effects. The disclosed methods are modular, biocompatible, and adaptable to a variety of gene editing contexts.

Dependencies are inferred from the claim text.