Determination of local chromatin interactions using a combined CRISPR and peroxidase APEX2 system
Determination of local chromatin interactions using a combined CRISPR and peroxidase APEX2 system
复制标题
使用 CRISPR 和过氧化物酶 APEX2 组合系统测定局部染色质相互作用
DOI：
10.1093/nar/gkz134
复制
发表时间：
2019-05-21
影响因子：
14.9
通讯作者：
Liu, Yun
中科院分区：
文献类型：
--
作者：
Qiu, Wenqing;Xu, Zhijiao;Liu, Yun
The architecture and function of chromatin are largely regulated by local interacting molecules, such as transcription factors and noncoding RNAs. However, our understanding of these regulatory molecules at a given locus is limited because of technical difficulties. Here, we describe the use of Clustered Regularly Interspaced Short Palindromic Repeats and an engineered ascorbate peroxidase 2 (APEX2) system to investigate local chromatin interactions (CAPLOCUS). We showed that with specific small-guide RNA targets, CAPLOCUS could efficiently identify both repetitive genomic regions and single-copy genomic locus with high resolution. Genome-wide sequencing revealed known and potential long-range chromatin interactions for a specific single-copy locus. CAPLOCUS also identified telomere-associated RNAs. CAPLOCUS, followed by mass spectrometry, identified both known and novel telomere-associated proteins in their native states. Thus, CAPLOCUS may be a useful approach for studying local interacting molecules at any given chromosomal location.
