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

By Bushra Raj, Daniel E. Wagner, Aaron McKenna, Shristi Pandey, Allon M. Klein, Jay Shendure, James A. Gagnon, Alexander F. Schier

Posted 19 Oct 2017
bioRxiv DOI: 10.1101/205534 (published DOI: 10.1038/nbt.4103)

Hundreds of cell types are generated during development, but their lineage relationships are largely elusive. Here we report a technology, scGESTALT, which combines cell type identification by single-cell RNA sequencing with lineage recording by cumulative barcode editing. We sequenced ~60,000 transcriptomes from the juvenile zebrafish brain and identified more than 100 cell types and marker genes. We engineered an inducible system that combines early and late barcode editing and isolated thousands of single-cell transcriptomes and their associated barcodes. The large diversity of edited barcodes and cell types enabled the generation of lineage trees with hundreds of branches. Inspection of lineage trajectories identified restrictions at the level of cell types and brain regions and helped uncover gene expression cascades during differentiation. These results establish scGESTALT as a new and widely applicable tool to simultaneously characterize the molecular identities and lineage histories of thousands of cells during development and disease.

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