Affiliation:
1. Key Laboratory of Organ Regeneration and Transplantation of Ministry of Education, Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, Jilin 130061, P.R. China
2. Stanford University Medical School, VA Palo Alto Health Care System, Palo Alto, CA 94304, USA
3. Department of Endocrinology, Xiangya Hospital, Central South University, Changsha, Hunan, P.R. China
Abstract
AbstractFormation of a pluripotency-specific chromatin network is a critical event in reprogramming somatic cells into pluripotent status. To characterize the regulatory components in this process, we used ‘chromatin RNA in situ reverse transcription sequencing’ (CRIST-seq) to profile RNA components that interact with the pluripotency master gene Oct4. Using this approach, we identified a novel nuclear lncRNA Oplr16 that was closely involved in the initiation of reprogramming. Oplr16 not only interacted with the Oct4 promoter and regulated its activity, but it was also specifically activated during reprogramming to pluripotency. Active expression of Oplr16 was required for optimal maintenance of pluripotency in embryonic stem cells. Oplr16 was also able to enhance reprogramming of fibroblasts into pluripotent cells. RNA reverse transcription-associated trap sequencing (RAT-seq) indicated that Oplr16 interacted with multiple target genes related to stem cell self-renewal. Of note, Oplr16 utilized its 3′-fragment to recruit the chromatin factor SMC1 to orchestrate pluripotency-specific intrachromosomal looping. After binding to the Oct4 promoter, Oplr16 recruited TET2 to induce DNA demethylation and activate Oct4 in fibroblasts, leading to enhanced reprogramming. These data suggest that Oplr16 may act as a pivotal chromatin factor to control stem cell fate by modulating chromatin architecture and DNA demethylation.
Funder
National Key Research and Development Program of China
National Natural Science Foundation of China
Ministry of Education
National Basic Research Program of China
Research on Chronic Noncommunicable Diseases Prevention
Control of National Ministry of Science and Technology
National Health Development Planning Commission Major Disease Prevention
Control of Science and Technology Plan of Action, Cancer Prevention and Control
Natural Science Foundation of Jilin Province
Provincial Science Fund of Jilin Province Development and Reform Commission
California Institute of Regenerative Medicine
Department of Veterans Affairs
Publisher
Oxford University Press (OUP)