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Red2Flpe-SCON: a versatile, multicolor strategy for generating mosaic conditional knockout mice

Szu-Hsien Sam Wu1,2, Somi Kim3, Heetak Lee4

  • 1Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna BioCenter (VBC), Dr. Bohr-Gasse 3, 1030, Vienna, Austria.

Nature Communications
|June 11, 2024
PubMed

Insights

A new Red2Flpe-SCON mouse model allows multicolor lineage tracing of mutant and wild-type cells. This system revealed that the Sox2 gene is crucial for stem cell proliferation and differentiation, not maintenance.

Area of Science:

  • Developmental Biology
  • Genetics
  • Stem Cell Biology

Background:

  • Image-based lineage tracing is vital for studying tissue dynamics and cell potential.
  • Previous Red2Onco models facilitated oncogenic clone analysis.
  • A need exists for systems that can track both mutant and wild-type cells simultaneously.

Purpose of the Study:

  • To develop a novel mosaic knockout system for multicolor lineage tracing.
  • To investigate the role of Sox2 in adult stem cell function using this new system.

Main Methods:

  • Development of the Red2Flpe mouse line for clone-specific Flpe expression.
  • Implementation of the FRT-based SCON (Short Conditional IntrON) method for tunable mosaic knockouts.
  • Application of the Red2Flpe-SCON system for Sox2 mutant clonal analysis in mouse esophageal epithelium.

Main Results:

  • The Red2Flpe-SCON system enables multicolor labeling of mutant and wild-type cells.
  • Sox2 mutant clonal analysis in the esophageal epithelium was successfully performed.
  • Sox2 was found to be less critical for stem cell maintenance but essential for proliferation and differentiation.

Conclusions:

  • The Red2Flpe-SCON system is a powerful tool for mosaic analysis in vivo.
  • Sox2 plays a critical role in regulating adult stem cell proliferation and differentiation.
  • This study refines our understanding of stem cell gene function in tissue homeostasis.

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