In vivo genome editing using Staphylococcus aureus Cas9
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Zhang_In vivo.pdf
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3.63 MB
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Author(s)
Ran, F. Ann
Cong, Le
Yan, Winston X.
Gootenberg, Jonathan S.
Kriz, Andrea J.
Zetsche, Bernd
Shalem, Ophir
Wu, Xuebing
Makarova, Kira S.
Koonin, Eugene V.
Date Issued
April 2015
Journal
Nature
Publisher
Nature Publishing Group
Citation
Ran, F. Ann, Le Cong, Winston X. Yan, David A. Scott, Jonathan S. Gootenberg, Andrea J. Kriz, Bernd Zetsche, et al. “In Vivo Genome Editing Using Staphylococcus Aureus Cas9.” Nature 520, no. 7546 (April 1, 2015): 186–91.
Version
Author's final manuscript
Abstract
The RNA-guided endonuclease Cas9 has emerged as a versatile genome-editing platform. However, the size of the commonly used Cas9 from Streptococcus pyogenes (SpCas9) limits its utility for basic research and therapeutic applications that use the highly versatile adeno-associated virus (AAV) delivery vehicle. Here, we characterize six smaller Cas9 orthologues and show that Cas9 from Staphylococcus aureus (SaCas9) can edit the genome with efficiencies similar to those of SpCas9, while being more than 1 kilobase shorter. We packaged SaCas9 and its single guide RNA expression cassette into a single AAV vector and targeted the cholesterol regulatory gene Pcsk9 in the mouse liver. Within one week of injection, we observed >40% gene modification, accompanied by significant reductions in serum Pcsk9 and total cholesterol levels. We further assess the genome-wide targeting specificity of SaCas9 and SpCas9 using BLESS, and demonstrate that SaCas9-mediated in vivo genome editing has the potential to be efficient and specific.
MIT Department
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
McGovern Institute for Brain Research at MIT
Koch Institute for Integrative Cancer Research at MIT
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/nature14299
