High-resolution analysis of DNA regulatory elements by synthetic saturation mutagenesis

被引：253

作者：

Patwardhan, Rupali P. ^{[1
]}

Lee, Choli ^{[1
]}

Litvin, Oren ^{[2
,3
]}

Young, David L. ^{[1
]}

Pe'er, Dana ^{[2
,3
]}

Shendure, Jay ^{[1
]}

机构：

[1] Univ Washington, Dept Genome Sci, Seattle, WA 98195 USA

[2] Columbia Univ, Dept Biol Sci, New York, NY 10027 USA

[3] Columbia Univ, Inst Computat Biol & Bioinformat, New York, NY USA

来源：

NATURE BIOTECHNOLOGY | 2009年 / 27卷 / 12期

关键词：

GENOME-WIDE ANALYSIS; GENE-EXPRESSION; PROMOTER;

D O I：

10.1038/nbt.1589

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：

We present a method that harnesses massively parallel DNA synthesis and sequencing for the high-throughput functional analysis of regulatory sequences at single-nucleotide resolution. As a proof of concept, we quantitatively assayed the effects of all possible single-nucleotide mutations for three bacteriophage promoters and three mammalian core promoters in a single experiment per promoter. The method may also serve as a rapid screening tool for regulatory element engineering in synthetic biology.

引用

页码：1173 / 1175

页数：3

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