Lyon Kenneth, Stasevich Timothy J
Department of Biochemistry and Molecular Biology and the Institute for Genome Architecture and Function, Colorado State University, Fort Collins, CO, USA.
Department of Biochemistry and Molecular Biology and the Institute for Genome Architecture and Function, Colorado State University, Fort Collins, CO, USA.
Trends Genet. 2017 May;33(5):322-335. doi: 10.1016/j.tig.2017.02.003. Epub 2017 Mar 27.
Antibody derivatives, such as antibody fragments (Fabs) and single-chain variable fragments (scFvs), are now being used to image traditionally hard-to-see protein subpopulations, including nascent polypeptides being translated and post-translationally modified proteins. This has allowed researchers to directly image and quantify, for the first time, translation initiation and elongation kinetics with single-transcript resolution and the temporal ordering and kinetics of post-translational histone and RNA polymerase II modifications. Here, we review these developments and discuss the strengths and weaknesses of live-cell imaging with antibody-based probes. Further development of these probes will increase their versatility and open new avenues of research for dissecting complex gene regulatory dynamics.
抗体衍生物，如抗体片段（Fabs）和单链可变片段（scFvs），目前正被用于对传统上难以观察到的蛋白质亚群进行成像，包括正在翻译的新生多肽和翻译后修饰的蛋白质。这使得研究人员首次能够以单转录本分辨率直接成像和量化翻译起始和延伸动力学，以及翻译后组蛋白和RNA聚合酶II修饰的时间顺序和动力学。在这里，我们回顾这些进展，并讨论基于抗体的探针进行活细胞成像的优缺点。这些探针的进一步发展将增加其通用性，并为剖析复杂的基因调控动力学开辟新的研究途径。
