测量酿酒酵母体内三核苷酸重复序列的动态行为。

Measuring Dynamic Behavior of Trinucleotide Repeat Tracts In Vivo in Saccharomyces cerevisiae.

作者信息

Williams Gregory M, Surtees Jennifer A

机构信息

Department of Biochemistry, School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY, 14214, USA.

Genetics, Genomics and Bioinformatics Program, Jacobs School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY, 14214, USA.

出版信息

Methods Mol Biol. 2018;1672:439-470. doi: 10.1007/978-1-4939-7306-4_30.

DOI:10.1007/978-1-4939-7306-4_30

PMID:29043641

Abstract

Trinucleotide repeat (TNR) tracts are inherently unstable during DNA replication, leading to repeat expansions and/or contractions. Expanded tracts are the cause of over 40 neurodegenerative and neuromuscular diseases. In this chapter, we focus on the (CNG) repeat sequences that, when expanded, lead to Huntington's disease (HD), myotonic dystrophy type 1 (DM1), and a number of other neurodegenerative diseases. We describe a series of in vivo assays, using the model system Saccharomyces cerevisiae, to determine and characterize the dynamic behavior of TNR tracts that are in the early stages of expansion, i.e., the so-called threshold range. Through a series of time courses and PCR-based assays, dynamic changes in tract length can be observed as a function of time. These assays can ultimately be used to determine how genetic factors influence the process of tract expansion in these early stages.

摘要

三核苷酸重复（TNR）序列在DNA复制过程中本质上是不稳定的，会导致重复序列的扩增和/或收缩。扩增的序列是40多种神经退行性疾病和神经肌肉疾病的病因。在本章中，我们重点关注（CNG）重复序列，当这些序列扩增时，会导致亨廷顿舞蹈病（HD）、1型强直性肌营养不良（DM1）以及许多其他神经退行性疾病。我们描述了一系列体内试验，利用酿酒酵母模型系统，来确定和表征处于扩增早期阶段（即所谓的阈值范围）的TNR序列的动态行为。通过一系列时间进程和基于聚合酶链式反应（PCR）的试验，可以观察到序列长度随时间的动态变化。这些试验最终可用于确定遗传因素如何在这些早期阶段影响序列扩增过程。