In vivo genome editing using Staphylococcus aureus Cas9

Details

• Alternative Title:
  Nature
• Personal Author:
  Ran, F. Ann ; Cong, Le ; Yan, Winston X. ; Scott, David A. ; Gootenberg, Jonathan S. ; Kriz, Andrea J. ; Zetsche, Bernd ; Shalem, Ophir ; Wu, Xuebing ; Makarova, Kira S. ; Koonin, Eugene ; Sharp, Phillip A. ; Zhang, Feng
• Description:
  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.
• Subjects:
  Animals Article Base Sequence Cholesterol CRISPR-Associated Proteins Gene Targeting Genetic Engineering Genome Liver Male Mice Mice, Inbred C57BL Proprotein Convertases Serine Endopeptidases Staphylococcus Aureus Substrate Specificity

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• Source:
  Nature. 520(7546):186-191.
• Pubmed ID:
  25830891
• Pubmed Central ID:
  PMC4393360
• Document Type:
  Journal Article
• Funding:
  5DP1-MH100706/DP/NCCDPHP CDC HHS/United States ; 5P30EY012196-17/EY/NEI NIH HHS/United States ; 5R01DK097768-03/DK/NIDDK NIH HHS/United States ; DP1 MH100706/MH/NIMH NIH HHS/United States ; P01-CA42063/CA/NCI NIH HHS/United States ; P30-CA14051/CA/NCI NIH HHS/United States ; R01-CA133404/CA/NCI NIH HHS/United States ; R01-GM34277/GM/NIGMS NIH HHS/United States ; T32 GM007753/GM/NIGMS NIH HHS/United States ; T32 GM008313/GM/NIGMS NIH HHS/United States ; T32GM007753/GM/NIGMS NIH HHS/United States ; Howard Hughes Medical Institute/United States
• Volume:
  520
• Issue:
  7546
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:3054f606101ba0aa1acef30506fd1fec26ce769e084270f78087c5fcd413eddd
• Download URL:
  https://stacks.cdc.gov/view/cdc/30379/cdc_30379_DS1.pdf
• File Type:
  [PDF - 3.63 MB ]