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Am J Respir Cell Mol Biol. 2020 Mar 24. doi: 10.1165/rcmb.2019-0416MA. [Epub ahead of print]

Phenotypic Diversity Caused by Differential Expression of SFTPC-Cre Transgenic Alleles.

Author information

1
University of Pittsburgh Department of Medicine, 199716, Pulmonary, Allergy, and Critical Care Medicine, Pittsburgh, Pennsylvania, United States.
2
University of Pittsburgh Department of Medicine, 199716, Pulmonary, Allergy, and Critical Care Medicine, Pittsburgh, Pennsylvania, United States; jalder@pitt.edu.

Abstract

Type II alveolar epithelial cells (AEC2s) play an essential role in the function and maintenance of the pulmonary epithelium. Several transgenic mice have been developed to study the function of these cells in vivo by using the human surfactant protein C (SFTPC) promoter to drive expression of Cre recombinase. The precise activity of each of these transgenic alleles has not been studied and previous reports suggest that their activity can depend on breeding strategies. We bred mice with a conditional allele of the essential telomere capping protein TRF2 with two different SFTPC-Cre transgenic strains and observed opposite phenotypes (100% lethality versus 100% viability). We characterized the Cre recombinase activity in these two transgenic lines and found that the contrasting phenotypes were driven by difference in embryonic expression of the two transgenes, likely due to position effects or differences in the transgenic constructs. We also tested if SFTPC-Cre activity was dependent on maternal or paternal inheritance. When paternally inherited, both SFTPC-Cre alleles produced offspring with constitutive reporter activity independent of the inheritance of the Cre allele, suggesting that Cre recombinase was expressed in the male germline prior to meiosis. Immunohistochemical analysis of the testis showed reporter activity during spermatogenesis. Analysis of single-cell RNA sequencing data from murine and human testis demonstrated SFPTC expression uniquely during human spermatogenesis, suggesting that use of the human promoter in these constructs is responsible for male germline activity. Our data highlight the importance of careful analysis of transgenic allele activity and identify an SFTPC-Cre allele that is useful for pan-epithelial targeting in the mouse.

KEYWORDS:

Lung development; Surfactant protein C; lineage tracing

PMID:
32208105
DOI:
10.1165/rcmb.2019-0416MA

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