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Nat Biotechnol. 2009 Dec;27(12):1173-5. doi: 10.1038/nbt.1589.

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

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  • 1Department of Genome Sciences, University of Washington, Seattle, Washington, USA. rpatward@u.washington.edu

Abstract

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.

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