A new protocol for single-cell RNA-seq reveals stochastic gene expression during lag phase in budding yeast
A new protocol for single-cell RNA-seq reveals stochastic gene expression during lag phase in budding yeast
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DOI：
10.7554/elife.55320
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发表时间：
2020-05-18
期刊：
影响因子：
7.7
通讯作者：
Verstrepen, Kevin J.
中科院分区：
文献类型：
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作者：
Jariani, Abbas;Vermeersch, Lieselotte;Verstrepen, Kevin J.
目前用于酵母细胞的单细胞RNA测序（scRNA-seq）的方法与可用于哺乳动物细胞的最新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10 x Genomics基于液滴的单细胞RNA测序技术，以允许分析酵母细胞。该协议，这是基于液滴内的细胞原生质球，产生一个数量级更高的吞吐量相比，现有的方法。经过广泛的验证的方法，我们证明了它的使用，通过研究的动力学的反应的同基因酵母菌群的碳源的转变，揭示了异质性和潜在的分子过程中，这种转变。我们描述的方法为研究酵母细胞以及其他具有可降解细胞壁的细胞开辟了新的途径。
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.