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cas9

2 papers tagged “cas9

BiologyNature · Apr 2016 Open access

Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage

Alexis C. Komor, Yongjoo B. Kim, Michael S. Packer, John A. Zuris and David R. Liu

This paper introduced cytosine base editing (CBE), a strategy that fuses a catalytically impaired Cas9 to a cytidine deaminase to directly convert C-G base pairs to T-A in genomic DNA without inducing double-strand breaks or requiring a donor template. The authors showed that within a programmable target window the deaminase converts cytosine to uracil, which is then read as thymine, and that inhibiting base-excision repair markedly improves editing efficiency. The approach achieved precise single-base correction in human and other mammalian cells.

BiologyScience · Aug 2012 Open access

A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity

Martin Jinek, Krzysztof Chylinski, Ines Fonfara, Michael Hauer, Jennifer A. Doudna and Emmanuelle Charpentier

This study demonstrated that the CRISPR-associated protein Cas9 from Streptococcus pyogenes is an RNA-guided DNA endonuclease whose target specificity is determined by a dual-RNA structure formed by a CRISPR RNA (crRNA) base-paired to a trans-activating crRNA (tracrRNA). The authors showed that Cas9 introduces site-specific double-strand breaks in target DNA, with its HNH domain cleaving the complementary strand and its RuvC-like domain cleaving the noncomplementary strand. Critically, they engineered the two guide RNAs into a single chimeric guide RNA that still directed sequence-specific cleavage, establishing the system as a programmable tool for genome editing.