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

Lukasz Swiech, Matthias Heidenreich, Abhishek Banerjee, Naomi Habib, Yinqing Li, John Trombetta, Mriganka Sur, Feng Zhang*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

604 Scopus citations


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.

Original languageAmerican English
Pages (from-to)102-106
Number of pages5
JournalNature Biotechnology
Issue number1
StatePublished - 1 Jan 2015
Externally publishedYes

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