Xu Chuanyun, Luo Liqun
Department of Biology and Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.
Biology Graduate Program, Stanford University, CA 94305, USA.
bioRxiv. 2024 Dec 4:2024.12.02.626507. doi: 10.1101/2024.12.02.626507.
In this protocol, we introduce a sparse driver system for cell-type specific single-cell labeling and manipulation in , enabling complete and simultaneous expression of multiple transgenes in the same cells. The system precisely controls expression probability and sparsity via mutant sites with reduced recombination efficiency and tunable FLP levels adjusted by heat-shock durations. We demonstrate that this generalizable toolkit enables tunable sparsity, multi-color staining, single-cell trans-synaptic tracing, single-cell manipulation, and analysis of cell-autonomous gene function. For details on the use and execution of this protocol, please refer to Xu et al. 2024.
在本方案中，我们引入了一种稀疏驱动系统，用于在[具体生物]中进行细胞类型特异性的单细胞标记和操作，能够在同一细胞中实现多个转基因的完整且同时表达。该系统通过具有降低重组效率的突变位点以及由热休克持续时间调节的可调FLP水平，精确控制表达概率和稀疏性。我们证明，这个可推广的工具包能够实现可调稀疏性、多色染色、单细胞跨突触追踪、单细胞操作以及细胞自主基因功能分析。有关本方案使用和执行的详细信息，请参考Xu等人（2024年）的研究。
