Ge Ming-Hai, Wang Wei, Wu Tai-Hong, Wen Xin, Al-Sheikh Umar, Chen Li-Li, Yin Sheng-Wu, Wu Jing-Jing, Huang Jia-Hao, He Qing-Qin, Liu Hui, Li Rong, Wang Ping-Zhou, Wu Zheng-Xing
Key Laboratory of Molecular Biophysics of Ministry of Education, Institute of Biophysics and Biochemistry, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.
iScience. 2020 Sep 15;23(10):101567. doi: 10.1016/j.isci.2020.101567. eCollection 2020 Oct 23.
Specific recording, labeling, and spatiotemporal manipulating neurons are essential for neuroscience research. In this study, we developed a tripartite spatiotemporal gene induction system in , which is based on the knockout of two transcriptional terminators (stops in short) by two different recombinases FLP and CRE. The recombinase sites (P and ) flanked stops after a ubiquitous promoter terminate transcription of target genes. FLP and CRE, induced by two promoters of overlapping expression, remove the stops (subsequent FLP/CRE-out). The system provides an "AND" gate strategy for specific gene expression in single types of cell(s). Combined with an inducible promoter or element, the system can control the spatiotemporal expression of genes in defined cell types, especially in cells or tissues lacking a specific promoter. This tripartite FLP/CRE-out gene expression system is a simple, labor- and cost-saving toolbox for cell type-specific and inducible gene expression in .
特异性记录、标记和时空操纵神经元对于神经科学研究至关重要。在本研究中，我们在[具体物种或细胞类型未提及]中开发了一种三方时空基因诱导系统，该系统基于两种不同的重组酶FLP和CRE敲除两个转录终止子（简称为“stops”）。在一个普遍存在的启动子之后，重组酶位点（P和[另一个位点未提及]）侧翼的“stops”终止靶基因的转录。由两个重叠表达的启动子诱导的FLP和CRE去除“stops”（随后是FLP/CRE-out）。该系统为单一类型细胞中的特异性基因表达提供了一种“与”门策略。与诱导型启动子或元件相结合，该系统可以控制特定细胞类型中基因的时空表达，特别是在缺乏特异性启动子的细胞或组织中。这种三方FLP/CRE-out基因表达系统是一种简单、节省劳力和成本的工具箱，用于[具体物种或细胞类型未提及]中细胞类型特异性和诱导型基因表达。
