High-throughput, image-based screening of pooled genetic-variant libraries

被引：40

作者：

Emanuel, George ^{[1
,2
,3
,4
]}

Moffitt, Jeffrey R. ^{[1
,3
,4
]}

Zhuang, Xiaowei ^{[1
,3
,4
]}

机构：

[1] Harvard Univ, Howard Hughes Med Inst, Cambridge, MA 02138 USA

[2] Harvard Univ, Grad Program Biophys, Cambridge, MA 02138 USA

[3] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA

[4] Harvard Univ, Dept Phys, Cambridge, MA 02138 USA

来源：

NATURE METHODS | 2017年 / 14卷 / 12期

关键词：

RED-FLUORESCENT PROTEINS; IN-SITU; SINGLE; PHOTOSTABILITY; MOLECULES; CIRCUITS; PROBES; SEQ; GFP;

D O I：

10.1038/nmeth.4495

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We report a high-throughput screening method that allows diverse genotypes and corresponding phenotypes to be imaged in individual cells. We achieve genotyping by introducing barcoded genetic variants into cells as pooled libraries and reading the barcodes out using massively multiplexed fluorescence in situ hybridization. To demonstrate the power of image-based pooled screening, we identified brighter and more photostable variants of the fluorescent protein YFAST among 60,000 variants.

引用

页码：1159 / +

页数：9

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