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.
The tetrameric structure of a casposase bound to DNA and its biochemical properties show how a transposase could have evolved to perform CRISPR-Cas spacer acquisition.
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.
The CRISPR/Cas9 system can be used with recombinant AAV6 donor delivery to facilitate simultaneous, targeted integration into multiple genetic loci in hematopoietic stem and progrenitor cells.
Gene knock-out of the omega-1 ribonuclease of Schistosoma mansoni eggs resulted in immunologically impaired phenotype, showcasing the novel application of CRISPR/Cas9 genome editing and utility for functional genomics in schistosomes.
The protein p53 negatively impacts the ability of a CRISPR screen to discriminate between essential and non-essential genes, hence, p53 status should be considered in these screens.
Divergent Cas9 enzymes direct site-specific single-stranded RNA cleavage, reducing infection by RNA phage in vivo and enabling programmable, PAM-independent repression of gene expression in bacteria.