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

被引：587

作者：

Swiech, Lukasz ^{[1
,2
,3
]}

Heidenreich, Matthias ^{[1
,2
,3
]}

Banerjee, Abhishek ^{[4
]}

Habib, Naomi ^{[1
,2
,3
]}

Li, Yinqing ^{[1
,2
,5
]}

Trombetta, John ^{[1
]}

Sur, Mriganka ^{[4
]}

Zhang, Feng ^{[1
,2
,3
]}

机构：

[1] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[2] MIT, McGovern Inst Brain Res, Dept Brain & Cognit Sci, Cambridge, MA 02139 USA

[3] MIT, McGovern Inst Brain Res, Dept Biol Engn, Cambridge, MA 02139 USA

[4] MIT, Picower Inst Learning & Memory, Dept Brain & Cognit Sci, Cambridge, MA 02139 USA

[5] MIT, Dept Elect Engn & Comp Sci, Cambridge, MA 02139 USA

来源：

NATURE BIOTECHNOLOGY | 2015年 / 33卷 / 01期

关键词：

CPG-BINDING PROTEIN-2; RETT-SYNDROME; DUAL-RNA; TRANSCRIPTION; MECP2; CAS9; EXPRESSION; REPRESSION; NEURONS; MODEL;

D O I：

10.1038/nbt.3055

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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) 1 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.

引用

页码：102 / U286

页数：9

共 42 条

[1]

Banerjee Abhishek, 2012, Front Psychiatry, V3, P34, DOI 10.3389/fpsyt.2012.00034

[2] DEVELOPMENTS IN NEURONAL CELL-CULTURE [J].

BANKER, G ;

GOSLIN, K .

NATURE, 1988, 336 (6195) :185-186

[3] Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system [J].

Bikard, David ;

Jiang, Wenyan ;

Samai, Poulami ;

Hochschild, Ann ;

Zhang, Feng ;

Marraffini, Luciano A. .

NUCLEIC ACIDS RESEARCH, 2013, 41 (15) :7429-7437

[4] MeCP2, a key contributor to neurological disease, activates and represses transcription [J].

Chahrour, Maria ;

Jung, Sung Yun ;

Shaw, Chad ;

Zhou, Xiaobo ;

Wong, Stephen T. C. ;

Qin, Jun ;

Zoghbi, Huda Y. .

SCIENCE, 2008, 320 (5880) :1224-1229

[5] The story of Rett syndrome: From clinic to neurobiology [J].

Chahrour, Maria ;

Zoghbi, Huda Y. .

NEURON, 2007, 56 (03) :422-437

[6] MeCP2 controls excitatory synaptic strength by regulating glutamatergic synapse number [J].

Chao, Hsiao-Tuan ;

Zoghbi, Huda Y. ;

Rosenmund, Christian .

NEURON, 2007, 56 (01) :58-65

[7] Deficiency of methyl-CpG binding protein-2 in CNS neurons results in a Rett-like phenotype in mice [J].

Chen, RZ ;

Akbarian, S ;

Tudor, M ;

Jaenisch, R .

NATURE GENETICS, 2001, 27 (03) :327-331

[8] Multiplex Genome Engineering Using CRISPR/Cas Systems [J].

Cong, Le ;

Ran, F. Ann ;

Cox, David ;

Lin, Shuailiang ;

Barretto, Robert ;

Habib, Naomi ;

Hsu, Patrick D. ;

Wu, Xuebing ;

Jiang, Wenyan ;

Marraffini, Luciano A. ;

Zhang, Feng .

SCIENCE, 2013, 339 (6121) :819-823

[9] Permanent Alteration of PCSK9 With In Vivo CRISPR-Cas9 Genome Editing [J].

Ding, Qiurong ;

Strong, Alanna ;

Patel, Kevin M. ;

Ng, Sze-Ling ;

Gosis, Bridget S. ;

Regan, Stephanie N. ;

Cowan, Chad A. ;

Rader, Daniel J. ;

Musunuru, Kiran .

CIRCULATION RESEARCH, 2014, 115 (05) :488-+

[10]

Esvelt KM, 2013, NAT METHODS, V10, P1116, DOI [10.1038/nmeth.2681, 10.1038/NMETH.2681]

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