Carolin Anders

TL;DR: A crystal structure of Streptococcus pyogenes Cas9 in complex with a single-molecule guide RNA and a target DNA containing a canonical 5′-NGG-3′ PAM is reported, revealing that the PAM motif resides in a base-paired DNA duplex.

TL;DR: To compare the architectures and domain organization of diverse Cas9 proteins, the atomic structures of Cas9 from Streptococcus pyogenes and Actinomyces naeslundii and AnaCas9 were determined by x-ray crystallography and three-dimensional reconstructions of apo-SpyCas9, SpyCas9:RNA, and SpyCas 9:RNA:DNA were obtained by negative-stain single-particle electron microscopy.

TL;DR: The results establish that optimally solubilized, in vitro assembled fluorescent Cas9-sgRNA RNPs provide a reproducible reagent for direct and scalable loss-of-function studies and applications beyond zebrafish experiments that require maximal DNA cutting efficiency in vivo.

TL;DR: In this article, crystal structures of the VQR, EQR, and VRER SpCas9 variants bound to target DNAs containing their preferred NAAG PAM sequences were reported.

TL;DR: CrispRVariants as discussed by the authors is a R-based toolkit that resolves and localizes individual mutant alleles with respect to the endonuclease cut site and provides insight toward effective guide and amplicon design as well as the mutagenic process.