A compact, versatile drug-induced splicing switch system with minimal background expression
Gene-switch techniques hold promising applications in contemporary genetics research, particularly in disease treatment and genetic engineering. Here, we developed a compact drug-induced splicing system that maintains low background using a human ubiquitin C (hUBC) promoter and optimized drug (LMI070) binding sequences based on the Xon switch system. To ensure precise subcellular localization of the protein of interest (POI), we inserted a 2A self-cleaving peptide between the extra N-terminal peptide and POI. This streamlined and optimized switch system, named miniXon2G, effectively regulated POIs in different subcellular localizations both in vitro and in vivo. Furthermore, miniXon2G could be integrated into endogenous gene loci, resulting in precise, reversible regulation of target genes by both endogenous regulators and drugs. Overall, these findings highlight the performance of miniXon2G in controlling protein expression with great potential for general applicability to diverse biological scenarios requiring precise and delicate regulation.
Authors
- Shuangpeng Li (ORCID: https://orcid.org/0000-0003-3118-0289)
- Qingjian Zou (ORCID: https://orcid.org/0000-0002-7729-6220)
- Jiahui Xi
- Xiaoqing Zhou (ORCID: https://orcid.org/0000-0002-5334-1701)
- Min Chen (ORCID: https://orcid.org/0000-0003-3294-9248)
- Jinling Wang (ORCID: https://orcid.org/0000-0002-2723-8989)
- Yingying Xiao (ORCID: https://orcid.org/0009-0008-9279-7489)
- Xuan Lu (ORCID: https://orcid.org/0009-0005-0688-0808)
- Hui Yuan
- Liangxue Lai (ORCID: https://orcid.org/0000-0002-5256-6802)
- Shuo Lin
- Yue Chi
- Chengcheng Tang (ORCID: https://orcid.org/0000-0001-8962-3359)
Institutions
- Guangzhou Institutes of Biomedicine and Health (CN)
- Jiangmen Wuyi Traditional Chinese Medicine Hospital (CN)
- Wuyi University (CN)
Publication Details
- Journal
- Cell Reports Methods
- Published
- 2024-09-01
- DOI
- https://doi.org/10.1016/j.crmeth.2024.100842
- Citations
- 4
- Primary Topic
- RNA and protein synthesis mechanisms
- Type
- article
- Field-Weighted Citation Impact
- 0.94
Funders
- National Natural Science Foundation of China
- Natural Science Foundation of Guangdong Province
- National Key Research and Development Program of China
- Science and Technology Planning Project of Guangdong Province
- National Key Research and Development Program of China Stem Cell and Translational Research