In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9

Details

• Alternative Title:
  Nat Biotechnol
• Personal Author:
  Swiech, Lukasz ; Heidenreich, Matthias ; Banerjee, Abhishek ; Habib, Naomi ; Li, Yinqing ; Trombetta, John ; Sur, Mriganka ; Zhang, Feng
• Description:
  Probing gene function in the mammalian brain can be greatly assisted with methods to manipulate the genome of neurons in vivo. The clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated endonuclease (Cas)9 from Streptococcus pyogenes (SpCas9) can be used to edit single or multiple genes in replicating eukaryotic cells, resulting in frame-shifting insertion/deletion (indel) mutations and subsequent protein depletion. Here, we delivered SpCas9 and guide RNAs using adeno-associated viral (AAV) vectors to target single (Mecp2) as well as multiple genes (Dnmt1, Dnmt3a and Dnmt3b) in the adult mouse brain in vivo. We characterized the effects of genome modifications in postmitotic neurons using biochemical, genetic, electrophysiological and behavioral readouts. Our results demonstrate that AAV-mediated SpCas9 genome editing can enable reverse genetic studies of gene function in the brain.
• Subjects:
  Animals Article Clustered Regularly Interspaced Short Palindromic Repeats Streptococcus Pyogenes
• Source:
  Nat Biotechnol. 2014; 33(1):102-106
• Pubmed ID:
  25326897
• Pubmed Central ID:
  PMC4492112
• Document Type:
  Journal Article
• Funding:
  5DP1-MH100706/DP/NCCDPHP CDC HHSUnited States/ ; R01 EY007023/EY/NEI NIH HHSUnited States/ ; R01EY007023/EY/NEI NIH HHSUnited States/ ; DP1 MH100706/MH/NIMH NIH HHSUnited States/ ; R01 NS073124/NS/NINDS NIH HHSUnited States/ ; 5R01-NS073124/NS/NINDS NIH HHSUnited States/ ; R01 MH085802/MH/NIMH NIH HHSUnited States/ ; R01MH085802/MH/NIMH NIH HHSUnited States/
• Volume:
  33
• Issue:
  1
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:0948cb6932e06215b7f47bcec2420028f6fb2bbef654ecddab6393db1654ab89
• Download URL:
  https://stacks.cdc.gov/view/cdc/32021/cdc_32021_DS1.pdf
• File Type:
  [PDF - 806.79 KB ]