Simultaneous single-cell profiling of lineages and cell types in the vertebrate brain.

Simultaneous single-cell profiling of lineages and cell types in the vertebrate brain.
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DOI:
10.1038/nbt.4103
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发表时间:
2018-06
影响因子:
46.9
通讯作者:
Schier AF
Schier AF
中科院分区:
工程技术1区
文献类型:
--
作者:
Raj B;Wagner DE;McKenna A;Pandey S;Klein AM;Shendure J;Gagnon JA;Schier AF

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在发育过程中产生的数百种细胞类型之间的谱系关系很难重建。最近的一种方法GESTALT使用CRISPR-Cas9条形码编辑进行大规模谱系追踪,但仅限于早期开发,无法识别细胞类型。在这里,我们提出了scGESTALT,它结合了GESTALT的谱系记录能力与单细胞RNA测序的细胞类型鉴定。该方法依赖于一个诱导系统,该系统能够在多个时间点编辑条形码,从而捕获发育后期的谱系信息。对来自幼年斑马鱼大脑的约60,000个转录组进行测序,鉴定出>100种细胞类型和标记基因。利用这些数据,我们生成了具有数百个分支的谱系树,这些分支有助于揭示分化过程中细胞类型、大脑区域和基因表达级联水平的限制。scGESTALT可以应用于其他多细胞生物,以同时表征发育和疾病期间数千个细胞的分子身份和谱系历史。
The lineage relationships among the hundreds of cell types generated during development are difficult to reconstruct. A recent method, GESTALT, used CRISPR-Cas9 barcode editing for large-scale lineage tracing, but was restricted to early development and did not identify cell types. Here we present scGESTALT, which combines the lineage recording capabilities of GESTALT with cell-type identification by single-cell RNA sequencing. The method relies on an inducible system that enables barcodes to be edited at multiple time points, capturing lineage information from later stages of development. Sequencing of ~60,000 transcriptomes from the juvenile zebrafish brain identifies >100 cell types and marker genes. Using these data, we generate lineage trees with hundreds of branches that help uncover restrictions at the level of cell types, brain regions, and gene expression cascades during differentiation. scGESTALT can be applied to other multicellular organisms to simultaneously characterize molecular identities and lineage histories of thousands of cells during development and disease.
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