In vivo interrogation of gene function in the mammalian brain using CRISPR-Cas9
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Sur_In vivo interrogation.pdf
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Author(s)
Swiech, Lukasz
Heidenreich, Matthias
Banerjee, Abhishek
Habib, Naomi
Li, Yinqing
Trombetta, John
Sur, Mriganka
Zhang, Feng
Date Issued
October 2014
Journal
Nature Biotechnology
Publisher
Nature Publishing Group
Citation
Swiech, Lukasz, Matthias Heidenreich, Abhishek Banerjee, Naomi Habib, Yinqing Li, John Trombetta, Mriganka Sur, and Feng Zhang. “In Vivo Interrogation of Gene Function in the Mammalian Brain Using CRISPR-Cas9.” Nature Biotechnology 33, no. 1 (October 19, 2014): 102–6.
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Author's final manuscript
Abstract
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.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
McGovern Institute for Brain Research at MIT
Picower Institute for Learning and Memory
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/nbt.3055
