The luciferase reporter system of the MMP12 endogenous promoter for investigating transcriptional regulation of the human MMP12 gene
Electron. j. biotechnol
;
43: 55-61, Jan. 2020. tab, ilus, graf
Artigo
em Inglês
| LILACS
| ID: biblio-1087522
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.
Texto completo:
DisponíveL
Índice:
LILACS (Américas)
Assunto principal:
Metaloproteinase 12 da Matriz
/
Sistemas CRISPR-Cas
/
Luciferases
Limite:
Humanos
Idioma:
Inglês
Revista:
Electron. j. biotechnol
Assunto da revista:
Biotecnologia
Ano de publicação:
2020
Tipo de documento:
Artigo
País de afiliação:
China
/
Estados Unidos
Instituição/País de afiliação:
Northwestern University Feinberg School of Medicine/US
/
Shaanxi Normal University/CN
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