Cyp19a1-Cre-EGFP: A placenta-specific Cre transgenic mouse model for targeted gene recombination in trophoblast cells

Shu-Min Pan; Nian-Kun Chen; Xue-Yuan Li; Xiao-Zhen Xie; Xiong Cao; Zhi-Jian Wang

doi:10.1016/j.placenta.2025.06.020

Cyp19a1-Cre-EGFP: A placenta-specific Cre transgenic mouse model for targeted gene recombination in trophoblast cells

Placenta. 2025 Jul 1:168:209-218. doi: 10.1016/j.placenta.2025.06.020. Online ahead of print.

Authors

Shu-Min Pan¹, Nian-Kun Chen², Xue-Yuan Li³, Xiao-Zhen Xie³, Xiong Cao⁴, Zhi-Jian Wang⁵

Affiliations

¹ Key Laboratory of Mental Health of the Ministry of Education, Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Guangdong Province Key Laboratory of Psychiatric Disorders, Department of Neurobiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China; Department of Obstetrics, Foshan Maternity & Child Healthcare Hospital, Foshan, 528000, Guangdong, China.
² Department of Obstetrics and Gynecology, The First Affiliated Hospital, Sun Yat-sen University, China.
³ Department of Obstetrics, Foshan Maternity & Child Healthcare Hospital, Foshan, 528000, Guangdong, China.
⁴ Key Laboratory of Mental Health of the Ministry of Education, Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Guangdong Province Key Laboratory of Psychiatric Disorders, Department of Neurobiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. Electronic address: caoxiong@smu.edu.cn.
⁵ Department of Obstetrics and Gynecology, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology, Guangdong-Hong Kong-Macao Greater Bay Area Higher Education Joint Laboratory of Maternal-Fetal Medicine, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, 510150, China. Electronic address: wzjnfyy@163.com.

PMID: 40618517
DOI: 10.1016/j.placenta.2025.06.020

Abstract

Objective: The placenta plays a critical role in fetal development, yet placenta-specific gene manipulation remains a challenge due to widespread gene expression across maternal and embryonic tissues. Here, we describe the generation and characterization of Cyp19a1-Cre-EGFP transgenic mice, a novel Cre-loxP model for placenta-specific gene recombination.

Methods: A Cyp19a1-Cre-EGFP construct was generated using the Cyp19a1 promoter, driving Cre recombinase expression with an enhanced green fluorescent protein (EGFP) reporter. Transgenic mice were produced via pronuclear microinjection and backcrossed to establish stable Cre-expressing lines. Tissue specificity of Cre-EGFP expression was assessed by confocal microscopy, immunofluorescence, and PCR genotyping at embryonic day 13.5 (E13.5). To validate Cre-mediated recombination, Cyp19a1-Cre^+/-EGFP mice were crossed with Nr3c1^loxP/loxP (glucocorticoid receptor floxed) mice, and gene knockdown efficiency was evaluated by immunostaining.

Results: Cyp19a1-Cre-EGFP mice exhibited exclusive Cre expression in placental trophoblast cells, with no detectable EGFP fluorescence in maternal tissues (ovary, amygdala) or fetal liver. Transgenic mice displayed normal growth, fertility, and placental morphology. Cre-mediated recombination resulted in significant knockdown of GR expression (P < 0.05), confirming the model's efficiency for placenta-specific gene deletion.

Conclusion: The Cyp19a1-Cre-EGFP mouse model provides a highly specific and efficient tool for placental gene manipulation. Its applications extend to studying placental development, pregnancy disorders, and fetal programming, with potential relevance for preclinical models of placental disease.

Keywords: Cre-loxP; Gene recombination; Placenta; Pregnancy complications; Transgenic mouse model; Trophoblast.