Samuel H. Sternberg

TL;DR: Understanding how small RNAs are used to find and destroy foreign nucleic acids will provide new insights into the diverse mechanisms of RNA-controlled genetic silencing systems.

TL;DR: It is shown that both binding and cleavage of DNA by Cas9–RNA require recognition of a short trinucleotide protospacer adjacent motif (PAM) and that PAM interactions trigger Cas9 catalytic activity.

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: A new hyper-accurate Cas9 variant (HypaCas9) is designed that demonstrates high genome-wide specificity without compromising on-target activity in human cells and offers a more comprehensive model to rationalize and modify the balance between target recognition and nuclease activation for precision genome editing.

TL;DR: It is shown that Cas9 binds with high affinity to single-stranded RNA targets matching the Cas9-associated guide RNA sequence when the PAM is presented in trans as a separate DNA oligonucleotide, revealing a fundamental connection between PAM binding and substrate selection by Cas9 and highlighting the utility of Cas9 for programmable transcript recognition without the need for tags.