Research Paper Volume 12, Issue 8 pp 7431—7447

Sirt1 is regulated by miR-135a and involved in DNA damage repair during mouse cellular reprogramming

Andy Chun Hang Chen 1, 2, *, , Qian Peng 2, *, , Sze Wan Fong 1, , William Shu Biu Yeung 1, 2, , Yin Lau Lee 1, 2, ,

  • 1 Department of Obstetrics and Gynaecology, The University of Hong Kong, Hong Kong SAR, China
  • 2 Shenzhen Key Laboratory of Fertility Regulation, The University of Hong Kong Shenzhen Hospital, Shenzhen, China
* Co-first authors

received: December 30, 2019 ; accepted: March 30, 2020 ; published: April 26, 2020 ;

https://doi.org/10.18632/aging.103090
How to Cite

Copyright © 2020 Chen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Sirt1 facilitates the reprogramming of mouse somatic cells into induced pluripotent stem cells (iPSCs). It is regulated by micro-RNA and reported to be a target of miR-135a. However, their relationship and roles on cellular reprogramming remain unknown. In this study, we found negative correlations between miR-135a and Sirt1 during mouse embryonic stem cells differentiation and mouse embryonic fibroblasts reprogramming. We further found that the reprogramming efficiency was reduced by the overexpression of miR-135a precursor but induced by the miR-135a inhibitor. Co-immunoprecipitation followed by mass spectrometry identified 21 SIRT1 interacting proteins including KU70 and WRN, which were highly enriched for DNA damage repair. In accordance, Sirt1 activator resveratrol reduced DNA damage during the reprogramming process. Wrn was regulated by miR-135a and resveratrol partly rescued the impaired reprogramming efficiency induced by Wrn knockdown. This study showed Sirt1, being partly regulated by miR-135a, bound proteins involved in DNA damage repair and enhanced the iPSCs production.

Abbreviations

DAVID: Database for Annotation, Visualization and Integrated Discovery; DOX: Doxycycline; DSBs: Double strand breaks; ESCs: Embryonic stem cells; HR: Homologous recombination; iPSCs: Induced pluripotent stem cells; KEGG: Kyoto Encyclopedia of Genes and Genomes; MEFs: Mouse embryonic fibroblasts; miRNAs: MicroRNAs; MOI: Multiplicity of infection; MRN: Mre11-Rad50-Nbs1; NHEJ: Nonhomologous end joining; RSV: Resveratrol; VPA: Valproic acid.