Department of Biophysics, Johns Hopkins University, Baltimore, Maryland, USA.
Department of Biophysics, Johns Hopkins University, Baltimore, Maryland, USA; Physics Frontier Center (Center for Physics of Living Cells), University of Illinois, Urbana, Illinois, USA.
J Biol Chem. 2021 Sep;297(3):101086. doi: 10.1016/j.jbc.2021.101086. Epub 2021 Aug 14.
Transcriptional regulation is one of the key steps in determining gene expression. Diverse single-molecule techniques have been applied to characterize the stepwise progression of transcription, yielding complementary results. These techniques include, but are not limited to, fluorescence-based microscopy with single or multiple colors, force measuring and manipulating microscopy using magnetic field or light, and atomic force microscopy. Here, we summarize and evaluate these current methodologies in studying and resolving individual steps in the transcription reaction, which encompasses RNA polymerase binding, initiation, elongation, mRNA production, and termination. We also describe the advantages and disadvantages of each method for studying transcription.
转录调控是决定基因表达的关键步骤之一。多种单分子技术已被应用于描述转录的逐步进展，得出了互补的结果。这些技术包括但不限于单或多色荧光显微镜、使用磁场或光的力测量和操纵显微镜以及原子力显微镜。在这里，我们总结和评估了这些当前的方法学，用于研究和解析转录反应中的各个步骤，包括 RNA 聚合酶结合、起始、延伸、mRNA 产生和终止。我们还描述了每种方法在研究转录方面的优缺点。
