Chen Huimin, Larson Daniel R
Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Genes Dev. 2016 Aug 15;30(16):1796-810. doi: 10.1101/gad.281725.116.
The production of a single mRNA is the result of many sequential steps, from docking of transcription factors to polymerase initiation, elongation, splicing, and, finally, termination. Much of our knowledge about the fundamentals of RNA synthesis and processing come from ensemble in vitro biochemical measurements. Single-molecule approaches are very much in this same reductionist tradition but offer exquisite sensitivity in space and time along with the ability to observe heterogeneous behavior and actually manipulate macromolecules. These techniques can also be applied in vivo, allowing one to address questions in living cells that were previously restricted to reconstituted systems. In this review, we examine the unique insights that single-molecule techniques have yielded on the mechanisms of gene expression.
单个信使核糖核酸（mRNA）的产生是许多连续步骤的结果，从转录因子与聚合酶的对接、起始、延伸、剪接，到最终的终止。我们对RNA合成与加工基本原理的许多认识都来自体外生化测量的整体研究。单分子方法非常符合这种还原论传统，但在空间和时间上具有极高的灵敏度，同时具备观察异质性行为以及实际操纵大分子的能力。这些技术也可应用于体内，使人们能够解决以前仅限于重组系统的活细胞中的问题。在本综述中，我们将探讨单分子技术在基因表达机制方面所产生的独特见解。
