Journal article

Publication year: 2021

Journal: Frontiers in Cell and Developmental Biology

Volume number: 9

ISSN: 2296-634X

Open access status: Gold

DOI Link: https://doi.org/10.3389/fcell.2021.671841

Publisher: Frontiers Media

Abstract: 
Fgf10 is a key gene during development, homeostasis and repair after injury. We previously reported a knock-in Fgf10(Cre-ERT2) line (with the Cre-ERT2 cassette inserted in frame with the start codon of exon 1), called thereafter Fgf10(Ki-v1), to target FGF10(Pos) cells. While this line allowed fairly efficient and specific labeling of FGF10(Pos) cells during the embryonic stage, it failed to target these cells after birth, particularly in the postnatal lung, which has been the focus of our research. We report here the generation and validation of a new knock-in Fgf10(Cre-ERT2) line (called thereafter Fgf10(Ki-v2)) with the insertion of the expression cassette in frame with the stop codon of exon 3. Fgf10(Ki-v2/+) heterozygous mice exhibited comparable Fgf10 expression levels to wild type animals. However, a mismatch between Fgf10 and Cre expression levels was observed in Fgf10(Ki-v2/+) lungs. In addition, lung and limb agenesis were observed in homozygous embryos suggesting a loss of Fgf10 functional allele in Fgf10(Ki-v2) mice. Bioinformatic analysis shows that the 3 ' UTR, where the Cre-ERT2 cassette is inserted, contains numerous putative transcription factor binding sites. By crossing this line with tdTomato reporter line, we demonstrated that tdTomato expression faithfully recapitulated Fgf10 expression during development. Importantly, Fgf10(Ki-v2) mouse is capable of significantly targeting FGF10(Pos) cells in the adult lung. Therefore, despite the aforementioned limitations, this new Fgf10(Ki-v2) line opens the way for future mechanistic experiments involving the postnatal lung.

Harvard Citation style: Chu, X., Taghizadeh, S., Vazquez-Armendariz, A., Herold, S., Chong, L., Chen, C., et al. (2021) Validation of a Novel Fgf10^Cre-ERT2 Knock-in Mouse Line Targeting FGF10^Pos Cells Postnatally, Frontiers in Cell and Developmental Biology, 9, Article 671841. https://doi.org/10.3389/fcell.2021.671841

APA Citation style: Chu, X., Taghizadeh, S., Vazquez-Armendariz, A., Herold, S., Chong, L., Chen, C., Zhang, J., El Agha, E., & Bellusci, S. (2021). Validation of a Novel Fgf10^Cre-ERT2 Knock-in Mouse Line Targeting FGF10^Pos Cells Postnatally. Frontiers in Cell and Developmental Biology. 9, Article 671841. https://doi.org/10.3389/fcell.2021.671841