International Journal of Molecular Sciences (Aug 2023)

Analysis of the Promoter Regions of gga-miR-31 and Its Regulation by RA and C-jun in Chicken

  • Yingjie Wang,
  • Ruihong Kong,
  • Ke Xie,
  • Cai Hu,
  • Zongyi Zhao,
  • Yuhui Wu,
  • Qisheng Zuo,
  • Bichun Li,
  • Yani Zhang

DOI
https://doi.org/10.3390/ijms241512516
Journal volume & issue
Vol. 24, no. 15
p. 12516

Abstract

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The role of gga-miR-31 in chicken germ cell differentiation and spermatogenesis is of significant importance. The transcriptional properties of gga-miR-31 are crucial in establishing the foundation for the formation of chicken spermatogonia stem cells and spermatogenesis. In this study, a series of recombinant vectors including varying lengths of the gga-miR-31 promoter were predicted and constructed. Through the utilization of the dual luciferase reporting system, the upstream −2180~0 bp region of gga-miR-31 was identified as its promoter region. Furthermore, it was predicted and confirmed that the activity of the gga-miR-31 promoter is increased by retinoic acid (RA). The binding of RA to the gga-miR-31 and Stra8 promoter regions was found to be competitive. Through the deletion of C-jun binding sites and the manipulation of C-jun expression levels, it was determined that C-jun inhibits the activity of the gga-miR-31 promoter. Furthermore, the combined treatment of C-jun and RA demonstrated that the positive regulatory effect of RA on the gga-miR-31 promoter is attenuated in the presence of high levels of C-jun. Overall, this study establishes a foundation for further investigation into the regulatory mechanisms of gga-miR-31 action, and provides a new avenue for inducing chicken embryonic stem cells (ESC) to differentiate into spermatogonial stem cells (SSC), and sperm formation.

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