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.
An optimized CRISPR-Cas9 system enables multiplexed genome engineering for evolving biomolecules and pathways from chromosomally integrated DNA libraries.
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.
Degradosome-associated nucleases PNPase and RNase J2 are required for type III CRISPR immunity against diverse nucleic acid invaders originating from plasmid and phage.
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.
If released in the wild, current CRISPR-based gene drive systems designed to alter populations could spread much farther than intended, despite the evolution of drive resistance.
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.