Laboratory for Systems Biology, VIB-KU Leuven Center for Microbiology, Leuven, Belgium.
Laboratory of Genetics and Genomics, CMPG, Department M2S, KU Leuven, Leuven, Belgium.
Elife. 2020 May 18;9:e55320. doi: 10.7554/eLife.55320.
Current methods for single-cell RNA sequencing (scRNA-seq) of yeast cells do not match the throughput and relative simplicity of the state-of-the-art techniques that are available for mammalian cells. In this study, we report how 10x Genomics' droplet-based single-cell RNA sequencing technology can be modified to allow analysis of yeast cells. The protocol, which is based on in-droplet spheroplasting of the cells, yields an order-of-magnitude higher throughput in comparison to existing methods. After extensive validation of the method, we demonstrate its use by studying the dynamics of the response of isogenic yeast populations to a shift in carbon source, revealing the heterogeneity and underlying molecular processes during this shift. The method we describe opens new avenues for studies focusing on yeast cells, as well as other cells with a degradable cell wall.
目前用于酵母细胞单细胞 RNA 测序（scRNA-seq）的方法与现有的哺乳动物细胞的最先进技术的通量和相对简单性不匹配。在这项研究中，我们报告了如何修改 10x Genomics 的基于液滴的单细胞 RNA 测序技术，以允许分析酵母细胞。该方案基于细胞在液滴中的球形体形成，与现有方法相比，通量提高了一个数量级。在对该方法进行了广泛的验证之后，我们通过研究同基因酵母群体对碳源变化的响应动力学来证明其用途，揭示了这种转变过程中的异质性和潜在的分子过程。我们描述的方法为专注于酵母细胞以及具有可降解细胞壁的其他细胞的研究开辟了新途径。
