Volume 27, No 4, Apr 2017
ISSN: 1001-0602
EISSN: 1748-7838 2018
impact factor 17.848*
(Clarivate Analytics, 2019)
Volume 27 Issue 4, April 2017: 483-504 | Open Access
ORIGINAL ARTICLES
Visualization of aging-associated chromatin alterations with an engineered TALE system
Ruotong Ren1,2,*, Liping Deng1,2,3,*, Yanhong Xue1,2,*, Keiichiro Suzuki4,*, Weiqi Zhang1,2, Yang Yu5, Jun Wu4, Liang Sun6, Xiaojun Gong7, Huiqin Luan1, Fan Yang8, Zhenyu Ju9, Xiaoqing Ren1, Si Wang1, Hong Tang7, Lingling Geng1, Weizhou Zhang10, Jian Li6, Jie Qiao5, Tao Xu1,2, Jing Qu2,3 and Guang-Hui Liu1,2,8,11
1National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
2University of Chinese Academy of Sciences, Beijing 100049, China
3State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
4Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
5Department of Gynecology and Obstetrics, Peking University Third Hospital, Beijing 100191, China
6The MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of Gerontology, Beijing 100730, China
7Department of Pediatrics, Beijing Shijitan Hospital Capital Medical University, Peking University Ninth School of Clinical Medicine, Beijing 100038, China
8Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou, Guangdong 510632, China
9Institute of Aging Research, Hangzhou Normal University School of Medicine, Hangzhou, Zhejiang 311121, China
10Department of Pathology, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
11Beijing Institute for Brain Disorders, Beijing 100069, China
Correspondence: Guang-Hui Liu, E-mail: ghliu@ibp.ac.cn; Jing Qu, E-mail: qujing@ioz.ac.cn; Tao Xu,(xutao@ibp.ac.cn)
Visualization of specific genomic loci in live cells is a prerequisite for the investigation of dynamic changes in chromatin architecture during diverse biological processes, such as cellular aging. However, current precision genomic imaging methods are hampered by the lack of fluorescent probes with high specificity and signal-to-noise contrast. We find that conventional transcription activator-like effectors (TALEs) tend to form protein aggregates, thereby compromising their performance in imaging applications. Through screening, we found that fusing thioredoxin with TALEs prevented aggregate formation, unlocking the full power of TALE-based genomic imaging. Using thioredoxin-fused TALEs (TTALEs), we achieved high-quality imaging at various genomic loci and observed aging-associated (epi) genomic alterations at telomeres and centromeres in human and mouse premature aging models. Importantly, we identified attrition of ribosomal DNA repeats as a molecular marker for human aging. Our study establishes a simple and robust imaging method for precisely monitoring chromatin dynamics in vitro and in vivo.
10.1038/cr.2017.18
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