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PLoS One. 2014 Aug 20;9(8):e105584. doi: 10.1371/journal.pone.0105584. eCollection 2014.

CRISPR/Cas9-mediated gene knock-down in post-mitotic neurons.

Author information

1
Department of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, United States of America.

Abstract

The prokaryotic adaptive immune system CRISPR/Cas9 has recently been adapted for genome editing in eukaryotic cells. This technique allows for sequence-specific induction of double-strand breaks in genomic DNA of individual cells, effectively resulting in knock-out of targeted genes. It thus promises to be an ideal candidate for application in neuroscience where constitutive genetic modifications are frequently either lethal or ineffective due to adaptive changes of the brain. Here we use CRISPR/Cas9 to knock-out Grin1, the gene encoding the obligatory NMDA receptor subunit protein GluN1, in a sparse population of mouse pyramidal neurons. Within this genetically mosaic tissue, manipulated cells lack synaptic current mediated by NMDA-type glutamate receptors consistent with complete knock-out of the targeted gene. Our results show the first proof-of-principle demonstration of CRISPR/Cas9-mediated knock-down in neurons in vivo, where it can be a useful tool to study the function of specific proteins in neuronal circuits.

PMID:
25140704
PMCID:
PMC4139396
DOI:
10.1371/journal.pone.0105584
[Indexed for MEDLINE]
Free PMC Article

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