Abstract
SummaryWnt signal transduction controls tissue morphogenesis, maintenance and regeneration in all multicellular animals. In mammals, the WNT/CTNNB1 (Wnt/β‐catenin) pathway controls cell proliferation and cell fate decisions before and after birth. It plays a critical role at multiple stages of embryonic development, but also governs stem cell maintenance and homeostasis in adult tissues. However, it remains challenging to monitor endogenous WNT/CTNNB1 signaling dynamics in vivo. Here, we report the generation and characterization of a new knock‐in mouse strain that doubles as a fluorescent reporter and lineage tracing driver for WNT/CTNNB1 responsive cells. We introduced a multi‐cistronic targeting cassette at the 3′ end of the universal WNT/CTNNB1 target gene Axin2. The resulting knock‐in allele expresses a bright fluorescent reporter (3xNLS‐SGFP2) and a doxycycline‐inducible driver for lineage tracing (rtTA3). We show that the Axin2 P2A‐rtTA3‐T2A‐3xNLS‐SGFP2 strain labels WNT/CTNNB1 responsive cells at multiple anatomical sites during different stages of embryonic and postnatal development. It faithfully reports the subtle and dynamic changes in physiological WNT/CTNNB1 signaling activity that occur in vivo. We expect this mouse strain to be a useful resource for biologists who want to track and trace the location and developmental fate of WNT/CTNNB1 responsive stem cells in different contexts.
Highlights
WNT/CTNNB1 (Wnt/β-catenin) signaling controls multiple stages of development in the mammalian embryo, ranging from primary axis formation (Liu et al, 1999) to mammary gland morphogenesis (ChuAnoeska Agatha Alida van de Moosdijk and Yorick Bernardus Cornelis van de Grift contributed to this work.et al, 2004)
We designed the reporter to serve as a direct readout of Axin2 activity and to be suitable for live cell imaging
To promote equimolar expression of AXIN2, rtTA3, and 3xNLS-SGFP2, self-cleaving 2A peptides were favored over IRES sequences (Goedhart et al, 2011)
Summary
Wnt signal transduction controls tissue morphogenesis, maintenance and regeneration in all multicellular animals. The WNT/CTNNB1 (Wnt/β-catenin) pathway controls cell proliferation and cell fate decisions before and after birth It plays a critical role at multiple stages of embryonic development, and governs stem cell maintenance and homeostasis in adult tissues. We show that the Axin2P2A-rtTA3-T2A-3xNLS-SGFP2 strain labels WNT/CTNNB1 responsive cells at multiple anatomical sites during different stages of embryonic and postnatal development It faithfully reports the subtle and dynamic changes in physiological WNT/CTNNB1 signaling activity that occur in vivo. We expect this mouse strain to be a useful resource for biologists who want to track and trace the location and developmental fate of WNT/CTNNB1 responsive stem cells in different contexts. KEYWORDS Axin, beta-catenin, doxycycline-inducible lineage tracing, GFP reporter mouse, organoids
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