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Related Concept Videos

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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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.

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|June 11, 2024
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Summary
This summary is machine-generated.

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.

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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.