An RNA interference screen uncovers a new molecule in stem cell self-renewal and long-term regeneration

Ting Chen, Evan Heller, Slobodan Beronja, Naoki Oshimori, Nicole Stokes, Elaine Fuchs

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Adult stem cells sustain tissue maintenance and regeneration throughout the lifetime of an animal. These cells often reside in specific signalling niches that orchestrate the stem cell's balancing act between quiescence and cell-cycle re-entry based on the demand for tissue regeneration. How stem cells maintain their capacity to replenish themselves after tissue regeneration is poorly understood. Here we use RNA-interference-based loss-of-function screening as a powerful approach to uncover transcriptional regulators that govern the self-renewal capacity and regenerative potential of stem cells. Hair follicle stem cells provide an ideal model. These cells have been purified and characterized from their native niche in vivo and, in contrast to their rapidly dividing progeny, they can be maintained and passaged long-term in vitro. Focusing on the nuclear proteins and/or transcription factors that are enriched in stem cells compared with their progeny, we screened ∼2,000 short hairpin RNAs for their effect on long-term, but not short-term, stem cell self-renewal in vitro. To address the physiological relevance of our findings, we selected one candidate that was uncovered in the screen: TBX1. This transcription factor is expressed in many tissues but has not been studied in the context of stem cell biology. By conditionally ablating Tbx1 in vivo, we showed that during homeostasis, tissue regeneration occurs normally but is markedly delayed. We then devised an in vivo assay for stem cell replenishment and found that when challenged with repetitive rounds of regeneration, the Tbx1-deficient stem cell niche becomes progressively depleted. Addressing the mechanism of TBX1 action, we discovered that TBX1 acts as an intrinsic rheostat of BMP signalling: it is a gatekeeper that governs the transition between stem cell quiescence and proliferation in hair follicles. Our results validate the RNA interference screen and underscore its power in unearthing new molecules that govern stem cell self-renewal and tissue-regenerative potential.

Original languageEnglish (US)
Pages (from-to)104-108
Number of pages5
JournalNature
Volume485
Issue number7396
DOIs
StatePublished - May 3 2012
Externally publishedYes

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RNA Interference
Regeneration
Stem Cells
Stem Cell Niche
Hair Follicle
Transcription Factors
Colony-Forming Units Assay
Adult Stem Cells
Nuclear Proteins
Small Interfering RNA
Cell Biology
Cell Self Renewal
Cell Cycle
Homeostasis
Maintenance
Cell Proliferation

ASJC Scopus subject areas

  • General

Cite this

An RNA interference screen uncovers a new molecule in stem cell self-renewal and long-term regeneration. / Chen, Ting; Heller, Evan; Beronja, Slobodan; Oshimori, Naoki; Stokes, Nicole; Fuchs, Elaine.

In: Nature, Vol. 485, No. 7396, 03.05.2012, p. 104-108.

Research output: Contribution to journalArticle

Chen, Ting ; Heller, Evan ; Beronja, Slobodan ; Oshimori, Naoki ; Stokes, Nicole ; Fuchs, Elaine. / An RNA interference screen uncovers a new molecule in stem cell self-renewal and long-term regeneration. In: Nature. 2012 ; Vol. 485, No. 7396. pp. 104-108.
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