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In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9
  • Published Date:
    Oct 19 2014
  • Source:
    Nat Biotechnol. 2014; 33(1):102-106.
Filetype[PDF-806.79 KB]


Details:
  • Pubmed ID:
    25326897
  • Pubmed Central ID:
    PMC4492112
  • 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.

  • Document Type:
  • Collection(s):
  • Funding:
    5DP1-MH100706/DP/NCCDPHP CDC HHS/United States
    5R01-NS073124/NS/NINDS NIH HHS/United States
    DP1 MH100706/MH/NIMH NIH HHS/United States
    R01EY007023/EY/NEI NIH HHS/United States
    R01MH085802/MH/NIMH NIH HHS/United States
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