• Log In
  • New issue alert
  • Submit a manuscript
  • Register
  • Home
  • About
  • Editorial Board
  • Search
  • Archives
  • Current
  • Forthcoming

Share

Article Panel


Vol 43 (2020)
»Table of Contents
Reading Tools
  • About the author
  • How to cite this article
  • Indexing metadata
  • Print version
  • Look up terms
  • Finding References
  • Review policy

Related items
  • Author's work


Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International.
The luciferase reporter system of the MMP12 endogenous promoter for investigating transcriptional regulation of the human MMP12 gene | Du | Electronic Journal of Biotechnology
doi:10.1016/j.ejbt.2019.12.003
Electronic Journal of Biotechnology, Vol 43 (2020)

The luciferase reporter system of the MMP12 endogenous promoter for investigating transcriptional regulation of the human MMP12 gene

Chunhua Du, Yanqiu Wu, Yurong Ju, Junli Zhao, Peiyan Yang, Qinwen Mao, Haibin Xia



Abstract

Background: Matrix metalloproteinase 12 (MMP12), a member of MMPs, can take lots of roles including extracellular matrix component degradation, viral infection, inflammation, tissue remodeling and tumorigenesis. To explore the transcriptional regulation of MMP12 gene, a sensitive luciferase reporter HEK293 cell line for endogenous MMP12 promoter was generated by CRISPR/Cas9 technology.

Results: The HEK293-MMP12-T2A-luciferase-KI cell line was successfully established by CRISPR/Cas9 technology. The sequencing results indicated that one allele of the genome was proven to have a site-directed insertion of luciferase gene and another allele of the genome was confirmed to have additional 48 bp insertion in this cell line. The cell line was further demonstrated to be a sensitive reporter of the endogenous MMP12 promoter by applying transcription factors STAT3, AP-1 and SP-1 to the cell line. The reporter cell line was then screened with bioactive small molecule library, and a small molecule Tanshinone I was found to significantly inhibit the transcriptional activity of MMP12 gene in HEK293-MMP12-T2A-luciferase-KI cell line by luciferase activity assay, which was further confirmed to inhibit the expression of MMP12 mRNA in wild-type HEK293 cells.

Conclusions: This novel luciferase knock-in reporter system will be helpful for investigating the transcriptional regulation of MMP12 gene and screening the drugs targeting MMP12 gene.




Full Text: | PDF | HTML

ISSN:  0717-3458

Contact: edbiotec@pucv.cl

Pontificia Universidad Católica de Valparaíso
Av. Brasil 2950, Valparaíso, Chile
Copyright © 1997- 2023 by Electronic Journal of Biotechnology