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

Biotechnology Letters
Zihang LiHaibin Xia

Abstract

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. 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. 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.

References

Sep 29, 2004·The Journal of Biological Chemistry·Christopher M AdamsJoseph L Goldstein
Jul 31, 2013·Biochemical and Biophysical Research Communications·Chikako ShibataKazuhiko Koike
Aug 19, 2015·Biochemical and Biophysical Research Communications·Gyun-Sik OhSeung-Whan Kim

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Methods Mentioned

BETA
PCR
transfection
electrophoresis
genotyping

Software Mentioned

GraphPad Prism

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