期刊
CHEMICAL COMMUNICATIONS
卷 57, 期 75, 页码 9594-9597出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/d1cc02439f
关键词
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资金
- National Natural Science Foundation of China [81772010]
- Young Talent Support Plan'' of Xi'an Jiaotong University
- Natural Science Foundation of Shaanxi Province [2021JM-540, 2019JQ-924]
- Key Breeding Program by Collaborative innovation center of green manufacturing technology for traditional Chinese medicine in Shaanxi province [2019XT-1-03, 2019XT-1-05]
The study presents the design of a reporter gene for real-time imaging of the splicing process in living subjects and successfully detects dynamic pre-mRNA splicing activity. This provides a valuable tool for high-throughput screening of splicing modulators, potentially accelerating the development of new drugs for the treatment of disordered splicing diseases.
Dynamic changes in intron sequences, with their loss and gain, are poorly detected due to the limited methods for the non-invasive monitoring of the pre-mRNA splicing process. Here, we describe the design of a two-step transcriptional activation (TSTA) reporter for the real-time imaging of the splicing process in living subjects. By taking advantage of the strong transactivating properties of the GAL4-VP16 fusion protein, which can target upstream activation sequence (UAS) elements to boost subsequent firefly luciferase reporter gene expression, we successfully and consistently detected the dynamic pre-mRNA splicing activity in response to exogenous splicing modulators in living cells and animals. Our findings provide a valuable tool for the high-throughput screening of splicing modulators, which could speed up the development of new drugs for the treatment of disordered splicing diseases.
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