During CRISPR adaptation, Cas4 forms a ternary complex with the Cas1-Cas2 spacer integration complex, an interaction that coordinates substrate hand-off following precise, PAM-dependent prespacer processing prior to integration.
Through novel, cell-specific CRISPR tools to disrupt molecular clock genes, it was revealed that circadian rhythms are coordinated through a network, rather than by the clock of 'master regulatory' neurons.
Integrated modeling of sgRNA positioning, chromatin accessibility, and sequence features enables accurate prediction of effective target sites for CRISPR-mediated transcriptional modulation and design of highly active libraries for genome-scale genetic screens.
CRISPR interference (CRISPRi), which uses small guide RNAs to target catalytically dead Cas9 protein to chromatin, disrupts existing transcription units and generates new sites for initiation and termination of transcription on both strands of DNA.
Cloning-free 3Cs technology is developed for the generation of sequence-bias-free covalently closed circular synthesized (3Cs) CRISPR/Cas gRNA libraries that can interrogate the coding and noncoding human genome.