title: Single-cell analysis of proxy reporter allele-marked epithelial cells establishes intestinal stem cell hierarchy.
abstract: The recent development of targeted murine reporter alleles as proxies for intestinal stem cell activity has led to significant advances in our understanding of somatic stem cell hierarchies and dynamics. Analysis of these reporters has led to a model in which an indispensable reserve stem cell at the top of the hierarchy (marked by Bmi1 and Hopx reporters) gives rise to active intestinal stem cells (marked by an Lgr5 reporter). Despite these advances, controversy exists regarding the specificity and fidelity with which these alleles distinguish intestinal stem cell populations. Here, we undertake a comprehensive comparison of widely used proxy reporters including both CreERT2 and EGFP cassettes targeted to the Lgr5, Bmi1, and Hopx loci. Single-cell transcriptional profiling of these populations and their progeny reveals that reserve and active intestinal stem cells are molecularly and functionally distinct, supporting a two-stem-cell model for intestinal self-renewal.
authors.affiliation: Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Ning
authors.initials: N
authors.lastname: Li
authors.affiliation: Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Program in Cell and Molecular Biology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Maryam
authors.initials: M
authors.lastname: Yousefi
authors.affiliation: Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Angela
authors.initials: A
authors.lastname: Nakauka-Ddamba
authors.affiliation: Department of Cell and Developmental Biology, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Rajan
authors.initials: R
authors.lastname: Jain
authors.affiliation: PENN Molecular Profiling Facility, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: John
authors.initials: J
authors.lastname: Tobias
authors.affiliation: Department of Cell and Developmental Biology, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Regenerative Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Program in Cell and Molecular Biology, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Epstein
authors.affiliation: Department of Statistics, The Wharton School, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Shane T
authors.initials: ST
authors.lastname: Jensen
authors.affiliation: Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Cell and Developmental Biology, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Regenerative Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Center for Molecular Studies in Digestive and Liver Diseases, University of Pennsylvania, Philadelphia, PA 19104, USA; Program in Cell and Molecular Biology, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: lengner@vet.upenn.edu.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Lengner
mesh_terms: D000465:Algorithms; D000483:Alleles; D000818:Animals; D019070:Cell Lineage / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D016000:Cluster Analysis; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods; D017930:Genes, Reporter / Q000235:genetics; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000166:cytology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D008957:Models, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059010:Single-Cell Analysis / Q000379:methods*; D013234:Stem Cells / Q000145:classification / Q000166:cytology / Q000378:metabolism*
chemical_list: D008164:Luminescent Proteins