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 language | English (US) |
|---|---|
| Pages (from-to) | 110-116 |
| Number of pages | 7 |
| Journal | Journal of Biotechnology |
| Volume | 219 |
| DOIs | |
| State | Published - Feb 10 2016 |
Funding
This work was supported by the Fundamental Research Funds for the Central Universities ( GK201301010 , GK201503041 ), the China Postdoctoral Science Foundation ( 2015M570809 ) and research grants to H.X. and K.C. from the National Natural Science Foundation of China (No. 81272543 , No. 8147177 2 and No. 31470058 ) and the Natural Science Foundation of Shaanxi Province , China (No. 2014JM4113 ).
Keywords
- Gene editing
- Homologous recombination
- Knock-in
- Sox2
- TALEN
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology
- Bioengineering
- Biotechnology