Orthogonal gene knock out and activation with a catalytically active Cas9 nuclease

Details

• Alternative Title:
  Nat Biotechnol
• Personal Author:
  Dahlman, James E. ; Abudayyeh, Omar O. ; Joung, Julia ; Gootenberg, Jonathan S. ; Zhang, Feng ; Konermann, Silvana
• Description:
  We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing double-stranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells.
• Subjects:
  Bacterial Proteins CRISPR-Associated Protein 9 Endonucleases Gene Knockout Techniques HEK293 Cells Humans RNA, Guide, CRISPR-Cas Systems
• Source:
  Nat Biotechnol. 33(11):1159-1161
• Pubmed ID:
  26436575
• Pubmed Central ID:
  PMC4747789
• Document Type:
  Journal Article
• Funding:
  DP1 MH100706/MH/NIMH NIH HHSUnited States/ ; 1DP1-MH100706/DP/NCCDPHP CDC HHSUnited States/
• Volume:
  33
• Issue:
  11
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:b45fdec8926b629610e174ea44e66a492e2e8d6670473033c37333b81ba9ef58
• Download URL:
  https://stacks.cdc.gov/view/cdc/37904/cdc_37904_DS1.pdf
• File Type:
  [PDF - 398.15 KB ]