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.

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

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

Farrerol promotes the efficiency of CRISPR/Cas9-mediated genome editing in both cells and embryos.

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.

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.

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.