High-resolution analysis of DNA regulatory elements by synthetic saturation mutagenesis.
High-resolution analysis of DNA regulatory elements by synthetic saturation mutagenesis.
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DOI：
10.1038/nbt.1589
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发表时间：
2009-12
影响因子：
46.9
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中科院分区：
文献类型：
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我们提出了一种方法，利用大规模并行DNA合成和测序的高通量功能分析的调节序列在单核苷酸分辨率。作为概念的证明，我们定量测定了所有可能的单核苷酸突变对三个噬菌体启动子和三个哺乳动物核心启动子的影响，每个启动子在一个单一的实验。该方法也可作为合成生物学中调控元件工程的快速筛选工具。
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.