使用综合合成寡核苷酸分析研究先驱因子活性及其与染色质重塑因子的协同作用。

Investigating pioneer factor activity and its coordination with chromatin remodelers using integrated synthetic oligo assay.

作者信息

Chen Hengye, Yan Chao, Dhasarathy Archana, Kladde Michael, Bai Lu

机构信息

Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA; Center for Eukaryotic Gene Regulation, The Pennsylvania State University, University Park, PA 16802, USA.

出版信息

STAR Protoc. 2023 Jun 7;4(2):102279. doi: 10.1016/j.xpro.2023.102279.

DOI:10.1016/j.xpro.2023.102279

PMID:37289591

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10323128/

Abstract

Chromatin accessibility is regulated by pioneer factors (PFs) and chromatin remodelers (CRs). Here, we present a protocol, based on integrated synthetic oligonucleotide libraries in yeast, to systematically interrogate the nucleosome-displacing activities of PFs and their coordination with CRs. We describe steps for designing oligonucleotide sequences, constructing yeast libraries, measuring nucleosome configurations, and data analyses. This approach potentially can be adapted for use in higher eukaryotes to investigate the activities of many types of chromatin-associated factors. For complete details on the use and execution of this protocol, please refer to Yan et al., and Chen et al..

摘要

染色质可及性受先驱因子（PFs）和染色质重塑因子（CRs）调控。在此，我们提出一种基于酵母中整合合成寡核苷酸文库的方案，用于系统探究PFs的核小体置换活性及其与CRs的协同作用。我们描述了寡核苷酸序列设计、酵母文库构建、核小体构象测量及数据分析的步骤。该方法有可能适用于高等真核生物，以研究多种类型染色质相关因子的活性。有关此方案使用和执行的完整详细信息，请参考Yan等人及Chen等人的文献。