CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes

被引：2806

作者：

Gilbert, Luke A. ^{[1
,2
,4
,5
]}

Larson, Matthew H. ^{[1
,2
,4
,5
]}

Morsut, Leonardo ^{[1
,2
]}

Liu, Zairan ^{[10
]}

Brar, Gloria A. ^{[1
,2
,4
,5
]}

Torres, Sandra E. ^{[1
,2
,4
,5
]}

Stern-Ginossar, Noam ^{[1
,2
,4
,5
]}

Brandman, Onn ^{[1
,2
,4
,5
]}

Whitehead, Evan H. ^{[1
,3
,4
]}

Doudna, Jennifer A. ^{[2
,4
,5
,6
,7
,8
,9
]}

Lim, Wendell A. ^{[1
,2
,3
,4
]}

Weissman, Jonathan S. ^{[1
,2
,3
,4
,5
]}

Qi, Lei S. ^{[1
,3
,4
]}

机构：

[1] Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94158 USA

[2] Univ Calif San Francisco, Howard Hughes Med Inst, San Francisco, CA 94158 USA

[3] Univ Calif San Francisco, UCSF Ctr Syst & Synthet Biol, San Francisco, CA 94158 USA

[4] Calif Inst Quantitat Biomed Res, San Francisco, CA 94158 USA

[5] Univ Calif Berkeley, Ctr RNA Syst Biol, Berkeley, CA 94720 USA

[6] Univ Calif Berkeley, Dept Mol & Cellular Biol, Berkeley, CA 94720 USA

[7] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA

[8] Univ Calif Berkeley, Dept Bioengn, Berkeley, CA 94720 USA

[9] Univ Calif Berkeley, Lawrence Berkeley Natl Lab, Phys Biosci Div, Berkeley, CA 94720 USA

[10] Peking Univ, Beijing 100871, Peoples R China

来源：

CELL | 2013年 / 154卷 / 02期

关键词：

GENE-EXPRESSION; REPRESSION; BACTERIA; ACTIVATION; CHROMATIN; REVEALS; GENOMES; SYSTEMS; CELLS;

D O I：

10.1016/j.cell.2013.06.044

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The genetic interrogation and reprogramming of cells requires methods for robust and precise targeting of genes for expression or repression. The CRISPR-associated catalytically inactive dCas9 protein offers a general platform for RNA-guided DNA targeting. Here, we show that fusion of dCas9 to effector domains with distinct regulatory functions enables stable and efficient transcriptional repression or activation in human and yeast cells, with the site of delivery determined solely by a coexpressed short guide (sg)RNA. Coupling of dCas9 to a transcriptional repressor domain can robustly silence expression of multiple endogenous genes. RNA-seq analysis indicates that CRISPR interference (CRISPRi)-mediated transcriptional repression is highly specific. Our results establish that the CRISPR system can be used as a modular and flexible DNA-binding platform for the recruitment of proteins to a target DNA sequence, revealing the potential of CRISPRi as a general tool for the precise regulation of gene expression in eukaryotic cells.

引用

页码：442 / 451

页数：10

共 46 条

[11] CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III [J].

Deltcheva, Elitza ;

Chylinski, Krzysztof ;

Sharma, Cynthia M. ;

Gonzales, Karine ;

Chao, Yanjie ;

Pirzada, Zaid A. ;

Eckert, Maria R. ;

Vogel, Joerg ;

Charpentier, Emmanuelle .

NATURE, 2011, 471 (7340) :602-+

[12]

DEUSCHLE U, 1995, MOL CELL BIOL, V15, P1907

[13] Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems [J].

DiCarlo, James E. ;

Norville, Julie E. ;

Mali, Prashant ;

Rios, Xavier ;

Aach, John ;

Church, George M. .

NUCLEIC ACIDS RESEARCH, 2013, 41 (07) :4336-4343

[14] Functional Identification of Optimized RNAi Triggers Using a Massively Parallel Sensor Assay [J].

Fellmann, Christof ;

Zuber, Johannes ;

McJunkin, Katherine ;

Chang, Kenneth ;

Malone, Colin D. ;

Dickins, Ross A. ;

Xu, Qikai ;

Hengartner, Michael O. ;

Elledge, Stephen J. ;

Hannon, Gregory J. ;

Lowe, Scott W. .

MOLECULAR CELL, 2011, 41 (06) :733-746

[15]

Fisher AL, 1996, MOL CELL BIOL, V16, P2670

[16]

Fu Y., 2013, NAT BIOTECHNOL

[17]

Gibson DG, 2009, NAT METHODS, V6, P343, DOI [10.1038/NMETH.1318, 10.1038/nmeth.1318]

[18] TIGHT CONTROL OF GENE-EXPRESSION IN MAMMALIAN-CELLS BY TETRACYCLINE-RESPONSIVE PROMOTERS [J].

GOSSEN, M ;

BUJARD, H .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (12) :5547-5551

[19]

Gratz SJ., 2013, Genetics

[20] KRAB-Zinc Finger Proteins and KAP1 Can Mediate Long-Range Transcriptional Repression through Heterochromatin Spreading [J].

Groner, Anna C. ;

Meylan, Sylvain ;

Ciuffi, Angela ;

Zangger, Nadine ;

Ambrosini, Giovanna ;

Denervaud, Nicolas ;

Bucher, Philipp ;

Trono, Didier .

PLOS GENETICS, 2010, 6 (03)

← 1 2 3 4 5 →