Enhanced Bacterial Immunity and Mammalian Genome Editing via RNA-Polymerase-Mediated Dislodging of Cas9 from Double-Strand DNA Breaks

• Published in: Molecular cell Vol. 71; no. 1; pp. 42 - 55.e8
• Main Authors: Clarke, Ryan, Heler, Robert, MacDougall, Matthew S., Yeo, Nan Cher, Chavez, Alejandro, Regan, Maureen, Hanakahi, Leslyn, Church, George M., Marraffini, Luciano A., Merrill, Bradley J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05.07.2018
• Subjects: Animals; Bacteria - genetics; Bacteria - metabolism; Bacteria - virology; Bacteriophages - genetics; Bacteriophages - metabolism; Cas9; Cell Line; CRISPR; CRISPR-Associated Protein 9 - genetics; CRISPR-Associated Protein 9 - metabolism; DNA Breaks, Double-Stranded; DNA Repair; Gene Editing; genome editing; Mice; Mouse Embryonic Stem Cells - metabolism; phage biology; RNA polymerase; strand bias; transcription; CRISPR; genome editing; strand bias; phage biology; transcription; Cas9; RNA polymerase; DNA repair

The ability to target the Cas9 nuclease to DNA sequences via Watson-Crick base pairing with a single guide RNA (sgRNA) has provided a dynamic tool for genome editing and an essential component of adaptive immune systems in bacteria. After generating a double-stranded break (DSB), Cas9 remains stably...