A novel luciferase knock-in reporter system for studying transcriptional regulation of the human Sox2 gene

Dan Xiao, Weifeng Zhang, Yan Li, Kuan Liu, Junli Zhao, Xiaohong Sun, Linlin Shan, Qinwen Mao, Haibin Xia*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Sox2 is an important transcriptional factor that has multiple functions in stem cell maintenance and tumorigenesis. To investigate the transcriptional regulation of the Sox2 gene, a luciferase knock-in reporter system was established in HEK293 cells by placing the luciferase gene in the genome under the control of the Sox2 gene promoter using a transcription activator-like effector nuclease (TALEN)-mediated genome editing technique. PCR and Southern blot results confirmed the site-specific integration of a single copy of the exogenous luciferase gene into the genome. To prove the reliability and sensitivity of this novel luciferase knock-in system, a CRISPR/Cas transcription activation system for the Sox2 gene was constructed and applied to the knock-in system. The results indicated that luciferase activity was directly correlated with the activity of the Sox2 endogenous promoter. This novel system will be a useful tool to study the transcriptional regulation of Sox2, and has great potential in medical and industrial applications.

Original languageEnglish (US)
Pages (from-to)110-116
Number of pages7
JournalJournal of Biotechnology
Volume219
DOIs
StatePublished - Feb 10 2016

Keywords

  • Gene editing
  • Homologous recombination
  • Knock-in
  • Sox2
  • TALEN

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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