Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds

被引：717

作者：

Zalatan, Jesse G. ^{[1
,2
]}

Lee, Michael E. ^{[5
,6
]}

Almeida, Ricardo ^{[1
,2
]}

Gilbert, Luke A. ^{[1
,2
,7
]}

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

La Russa, Marie ^{[1
,3
,4
]}

Tsai, Jordan C. ^{[1
,2
]}

Weissman, Jonathan S. ^{[1
,2
,7
,8
]}

Dueber, John E. ^{[5
,6
]}

Qi, Lei S. ^{[1
,3
,8
]}

Lim, Wendell A. ^{[1
,2
,3
,8
]}

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, Ctr Syst & Synthet Biol, San Francisco, CA 94158 USA

[4] Univ Calif San Francisco, Biomed Sci Grad Program, San Francisco, CA 94158 USA

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

[6] Univ Calif Berkeley, Energy Biosci Inst, Berkeley, CA 94720 USA

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

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

来源：

CELL | 2015年 / 160卷 / 1-2期

关键词：

SACCHAROMYCES-CEREVISIAE; COAT PROTEIN; CAS9; COMBINATORIAL; OPTIMIZATION; REPRESSION; PATHWAY; SPECIFICITY; AFFINITY; CELLS;

D O I：

10.1016/j.cell.2014.11.052

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：

Eukaryotic cells execute complex transcriptional programs in which specific loci throughout the genome are regulated in distinct ways by targeted regulatory assemblies. We have applied this principle to generate synthetic CRISPR-based transcriptional programs in yeast and human cells. By extending guide RNAs to include effector protein recruitment sites, we construct modular scaffold RNAs that encode both target locus and regulatory action. Sets of scaffold RNAs can be used to generate synthetic multigene transcriptional programs in which some genes are activated and others are repressed. We apply this approach to flexibly redirect flux through a complex branched metabolic pathway in yeast. Moreover, these programs can be executed by inducing expression of the dCas9 protein, which acts as a single master regulatory control point. CRISPR-associated RNA scaffolds provide a powerful way to construct synthetic gene expression programs for a wide range of applications, including rewiring cell fates or engineering metabolic pathways.

引用

页码：339 / 350

页数：12

共 42 条

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