Functional assays for transcription mechanisms in high-throughput.
Functional assays for transcription mechanisms in high-throughput.
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DOI：
10.1016/j.ymeth.2019.02.017
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发表时间：
2019-04
期刊：
影响因子：
4.8
通讯作者：
C. Qiu;Craig D. Kaplan
中科院分区：
文献类型：
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作者：
C. Qiu;Craig D. Kaplan
核酸文库程序合成规模的急剧增加与深度测序相结合，推动了对核酸和蛋白质生物学的理解的进步。以核酸或编码蛋白质为中心的生物系统极大地受益于此类高通量研究，因为可以合成大型 DNA 变体池，并且可以通过深度测序轻松分析 DNA 或 RNA 转录产物。在这里，我们回顾了一般核酸和蛋白质研究的各种高通量功能测定的范围，然后以这些类型的研究如何深入了解 RNA 聚合酶 II (Pol II) 活性位点为例进行了讨论。我们讨论了这些实验的设计和执行中的方法论考虑因素，这对于任何系统的研究都应该有价值。
Dramatic increases in the scale of programmed synthesis of nucleic acid libraries coupled with deep sequencing have powered advances in understanding nucleic acid and protein biology. Biological systems centering on nucleic acids or encoded proteins greatly benefit from such high-throughput studies, given that large DNA variant pools can be synthesized and DNA, or RNA products of transcription, can be easily analyzed by deep sequencing. Here we review the scope of various high-throughput functional assays for studies of nucleic acids and proteins in general, followed by discussion of how these types of study have yielded insights into the RNA Polymerase II (Pol II) active site as an example. We discuss methodological considerations in the design and execution of these experiments that should be valuable to studies in any system.