De novo construction of mammalian synthetic inhibitory transcription factor and promoter pairs.
- Author:
Zijie YANG
1
;
Yijie PAN
1
;
Yiming CAI
1
;
Tong FU
1
;
Ao FENG
1
;
Yan LIU
1
;
Yiheng WANG
1
;
Xinxuan XIONG
1
;
Liang CAI
1
Author Information
- Publication Type:Journal Article
- Keywords: Bio-Brick; eukaryotic transcription regulation; synthetic biology; synthetic promoter; synthetic transcription factor
- MeSH: Animals; Gene Expression Regulation; Gene Regulatory Networks; Mammals; Promoter Regions, Genetic; Transcription Factors; Transcription, Genetic
- From: Chinese Journal of Biotechnology 2018;34(12):1886-1894
- CountryChina
- Language:Chinese
- Abstract: Transcriptional regulation is crucial for regulated gene expression. Due to the complexity, it has been difficult to engineer eukaryotic transcription factor (TF) and promoter pairs. The few availabilities of eukaryotic TF and promotor pairs limit their practical use for clinical or industrial applications. Here, we report a de novo construction of synthetic inhibitory transcription factor and promoter pairs for mammalian transcriptional regulation. The design of synthetic TF was based on the fusion of DNA binding domain and Kruppel associated box transcription regulating domain (KRAB). The synthetic promoter was constructed by inserting the corresponding TF response element after SV40 promoter. We constructed and tested five synthetic inhibitory transcription factor and promoter pairs in cultured mammalian cells. The inhibition capability and orthogonality were verified by flow cytometry. In summary, we demonstrate the feasibility of constructing mammalian inhibitory TF and promoter pairs, which could be standardized for advanced gene-circuit design and various applications in the mammalian synthetic biology.
