建立用于研究单细胞分辨率下组蛋白遗传模式的小鼠胚胎干细胞的方案。
Protocol for Establishing Mouse Embryonic Stem Cells to Study Histone Inheritance Pattern at Single-Cell Resolution.
机构信息
Department of Biology, The Johns Hopkins University, Baltimore, MD 21218, USA.
Centre for Stem Cells and Regenerative Medicine, King's College London, London SE1 9RT, UK.
出版信息
STAR Protoc. 2020 Nov 18;1(3):100178. doi: 10.1016/j.xpro.2020.100178. eCollection 2020 Dec 18.
Asymmetric histone inheritance can regulate cell-fate determination in male germline stem cells. However, it remains elusive how this mechanism may be used in mammalian system. Recently, we show mouse embryonic stem cells (mESCs) with Wnt3a beads display non-overlapping H3/H4 patterns. Here, we present a detailed protocol for tracking histone inheritance in asymmetrically dividing mESCs at single-cell resolution. This protocol will establish a new system to study histone inheritance in cultured mammalian cells and could be applied to other parallel systems. For complete details on the use and execution of this protocol, please refer to Tran et al. (2012), Habib et al. (2013), Lowndes et al. (2017), and Ma et al. (2020).
不对称的组蛋白遗传可以调节雄性生殖干细胞的细胞命运决定。然而，哺乳动物系统中如何利用这种机制仍不清楚。最近，我们发现用 Wnt3a 珠处理的小鼠胚胎干细胞 (mESC) 显示出非重叠的 H3/H4 模式。在这里，我们提供了一个详细的方案，用于在单细胞分辨率下跟踪不对称分裂的 mESC 中的组蛋白遗传。该方案将建立一个研究培养的哺乳动物细胞中组蛋白遗传的新系统，并可应用于其他平行系统。有关此方案的使用和执行的完整详细信息，请参阅 Tran 等人（2012 年）、Habib 等人（2013 年）、Lowndes 等人（2017 年）和 Ma 等人（2020 年）。
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