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

Author:
Li, Zihang, Zhao, Junli, Muhammad, Niaz, Wang, Dongyang, Mao, Qinwen, Xia, Haibin
Source:
Biotechnology letters 2018 v.40 no.11-12 pp. 1495-1506
ISSN:
0141-5492
Subject:
alleles, cell lines, drugs, enzyme activity, gene expression, humans, luciferase, polymerase chain reaction, screening, transcription (genetics)
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.
Agid:
6195068