笑对人生
(2022-04-30 21:43):
#paper The single-cell transcriptional landscape of mammalian organogenesis Cao J, et al. Nature. 2019 Feb;566(7745):496-502. doi: 10.1038/s41586-019-0969-x
该文章是利用一种名为sci-RNA-seq3超高通量单细胞测序技术,该技术一次实验可完成大约200万个细胞转录组测序建库,单人完成的时间为1周,成本为每个细胞0.01美元。该研究主要是对小鼠不同发育阶段的61个胚胎(E9.5到E13.5)的单细胞转录图谱进行了描述,该图谱命名为MOCA(mouse organogenesis cell altas)。文章虽然不是很新,但这是monocle3首次在scRNAseq(单细胞转录组测序)的应用案例,提供monocle3详尽的基本原理和分析思路。从文章的作者列表来看,也发现有monocle3软件开发者的名字。monocle3是一款用于scRNAseq拟时序分析的工具,为monocle2更新版本。虽然,monocle2是目前已发表文章的应用较为广泛的一款版本,但是它在实际使用时存在一些问题,第一,monocle2使用的细胞降维方式与seurat(一款流行的,能独立完成从细胞-基因表达矩阵到细胞降维聚类分群的scRNAseq工具)并不兼容;第二,该版本已被作者弃用并停止维护,实际应用中发现一些bug,却难以找到解决的方案。在生信分析中,如何选择软件往往是一个难题(这可能也是很多评测文章出现的原因)。作为一名工具的使用者,可以在充分理解算法原理的基础上,结合自己的研究,并通过调试,最终做出适当的选择。
The single-cell transcriptional landscape of mammalian organogenesis
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Abstract:
Mammalian organogenesis is a remarkable process. Within a short timeframe, the cells of the three germ layers transform into an embryo that includes most of the major internal and external organs. Here we investigate the transcriptional dynamics of mouse organogenesis at single-cell resolution. Using single-cell combinatorial indexing, we profiled the transcriptomes of around 2 million cells derived from 61 embryos staged between 9.5 and 13.5 days of gestation, in a single experiment. The resulting 'mouse organogenesis cell atlas' (MOCA) provides a global view of developmental processes during this critical window. We use Monocle 3 to identify hundreds of cell types and 56 trajectories, many of which are detected only because of the depth of cellular coverage, and collectively define thousands of corresponding marker genes. We explore the dynamics of gene expression within cell types and trajectories over time, including focused analyses of the apical ectodermal ridge, limb mesenchyme and skeletal muscle.
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