Format

Send to

Choose Destination
Nature. 2018 Dec;564(7735):219-224. doi: 10.1038/s41586-018-0744-4. Epub 2018 Dec 5.

Single-cell mapping of lineage and identity in direct reprogramming.

Biddy BA1,2,3, Kong W1,2,3, Kamimoto K1,2,3, Guo C1,2,3, Waye SE1,2,3, Sun T1,2,3,4, Morris SA5,6,7.

Author information

1
Department of Developmental Biology, Washington University School of Medicine in St Louis, St Louis, MO, USA.
2
Department of Genetics, Washington University School of Medicine in St Louis, St Louis, MO, USA.
3
Center of Regenerative Medicine, Washington University School of Medicine in St Louis, St Louis, MO, USA.
4
Nanomedicine Research Center, Department of Neurosurgery, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
5
Department of Developmental Biology, Washington University School of Medicine in St Louis, St Louis, MO, USA. s.morris@wustl.edu.
6
Department of Genetics, Washington University School of Medicine in St Louis, St Louis, MO, USA. s.morris@wustl.edu.
7
Center of Regenerative Medicine, Washington University School of Medicine in St Louis, St Louis, MO, USA. s.morris@wustl.edu.

Abstract

Direct lineage reprogramming involves the conversion of cellular identity. Single-cell technologies are useful for deconstructing the considerable heterogeneity that emerges during lineage conversion. However, lineage relationships are typically lost during cell processing, complicating trajectory reconstruction. Here we present 'CellTagging', a combinatorial cell-indexing methodology that enables parallel capture of clonal history and cell identity, in which sequential rounds of cell labelling enable the construction of multi-level lineage trees. CellTagging and longitudinal tracking of fibroblast to induced endoderm progenitor reprogramming reveals two distinct trajectories: one leading to successfully reprogrammed cells, and one leading to a 'dead-end' state, paths determined in the earliest stages of lineage conversion. We find that expression of a putative methyltransferase, Mettl7a1, is associated with the successful reprogramming trajectory; adding Mettl7a1 to the reprogramming cocktail increases the yield of induced endoderm progenitors. Together, these results demonstrate the utility of our lineage-tracing method for revealing the dynamics of direct reprogramming.

Comment in

PMID:
30518857
PMCID:
PMC6635140
DOI:
10.1038/s41586-018-0744-4
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center