Department of Anatomy, University of California, San Francisco, San Francisco, CA 94158, USA; Tetrad Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA.
Department of Anesthesiology & Pain Medicine, University of Washington, Seattle, WA 98195, USA.
STAR Protoc. 2022 Aug 18;3(3):101625. doi: 10.1016/j.xpro.2022.101625. eCollection 2022 Sep 16.
Existing techniques for transcriptional profiling of projection neurons could be applied to only one neuronal population per experiment. To increase throughput, we developed VECTORseq, which repurposes retrogradely infecting viruses to deliver multiplexable RNA barcodes, enabling projection anatomy to be read out in single-cell datasets. In this protocol, we describe the delivery of viral barcodes to mouse brain to label different projection neurons. We then detail single-cell or nuclei isolation for sequencing, followed by the analysis of single-cell sequencing data. For complete details on the use and execution of this protocol, please refer to Cheung et al. (2021).
现有的投射神经元转录组分析技术，每次实验只能针对一个神经元群体。为了提高通量，我们开发了 VECTORseq，它重新利用逆行感染病毒来递呈可多重编码的 RNA 条码，从而可以在单细胞数据集读取投射神经元的解剖结构。在本方案中，我们描述了将病毒条码递送到小鼠大脑中，以标记不同的投射神经元。然后详细介绍了单细胞或细胞核分离进行测序，随后对单细胞测序数据进行分析。有关此方案使用和执行的完整详细信息，请参阅 Cheung 等人（2021 年）。
