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Updated: Jun 18, 2026
A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates
Published on: August 21, 2018
High-resolution analysis of DNA regulatory elements by synthetic saturation mutagenesis
Rupali P Patwardhan1, Choli Lee, Oren Litvin
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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