Establishment of a HEK293 cell line by CRISPR/Cas9-mediated luciferase knock-in to study transcriptional regulation of the human SREBP1 gene

Zihang Li, Junli Zhao, Niaz Muhammad, Dongyang Wang, Qinwen Mao, Haibin Xia*

*Corresponding author for this work

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Objectives: To establish a HEK293 cell line with a luciferase knock-in reporter controlled by the endogenous SREBP1 promoter for investigating transcriptional regulation of the SREBP1 gene. Results: PCR confirmed the site-specific integration of a single copy of the exogenous luciferase gene into one allele of the genome and a 14 bp deletion of the targeted sequence in the other. Luciferase activity was directly correlated with the promoter activity of the endogenous SREBP1 gene in the HEK293-SREBP1-T2A-luciferase-KI cell line cell line. Conclusions: We successfully generated a novel luciferase knock-in reporter system, which will be very useful for studying transcriptional regulation of the SREBP1 gene and for screening drugs or chemical molecules that regulate SREBP1 gene expression.

Original languageEnglish (US)
JournalBiotechnology Letters
DOIs
StateAccepted/In press - Jan 1 2018

Keywords

  • CRISPR/Cas9
  • Cell line
  • Knock-in
  • SREBP1
  • Transcriptional regulation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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