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.

The inhibitory influence of nucleosomes on CRISPR-Cas9 is mitigated by nucleosomal DNA breathing and chromatin remodeling.

The guide RNA and donor DNA of the CRISPR/Cas system tolerate large chemical modifications and can be engineered for enhanced delivery and gene editing.

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.

Linking the DNA repair template to the Cas9 ribonucleoprotein complex enhances homology-directed repair of the induced DNA double strand break.

Building on previous work (Jinek et al., 2013), we report a simple and robust system to achieve high fidelity and high efficiency (30% of homologous recombination) genome engineering by homology-directed repair pathway in human cells using cell cycle synchronization and timed delivery of Cas9 ribonucleoprotein complexes.

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.

Conflicts between CRISPR-Cas systems and antibiotic resistance plasmids can be exploited to selectively eliminate antibiotic resistance from Enterococcus faecalis populations.