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Generation and identification of renal tubular epithelial cell C/EBPα conditional knockout mice

YING Ji, LIU Jian, ZHANG Li-wen, ZHONG Fang, ZHOU Fang-fang, WANG Wei-ming   

  1. Department of Nephrology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2016-10-28 Published:2016-11-29
  • Supported by:

    National Natural Science Foundation of China, 81270782， 30771000; National Basic Research Program of China, 2012CB517701, 2012CB517604; the National Science & Technology Pillar Program during the Twelfth Fiveyear Plan Period, 2011BAI10B00, 2011BAI10B06; Key Discipline Construction Project of Shanghai Municipal Commission of Health and Family Planning; Health Industry Specific Research Project, 201002010; Comparative Research, Scientific Research Project of Shanghai, 15140902800

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Abstract:

Objective·To generate renal tubular epithelial cell C/EBPα conditional knockout  mice, perform phenotypic identification, and lay a foundation for further study on the role of C/EBPα in renal interstitial fibrosis.Methods·C/EBPαflox/flox and pepck-Cre+ transgenic mice were bred and hybridized.The first generation offspring were mated to reproduce the second generation. Mice with C/EBPαflox/flox.pepck-Cre+ genotype (C/EBPα-/- mice) were identified using PCR. The mRNA and protein expressions of C/EBPα in renal tissues were measured using RT-qPCR and immunohistochemistry staining. The morphologic changes in renal tissues of C/EBPα-/- mice and wide type (WT) mice aged 6-8 weeks were observed using H-E staining and Masson staining.Results·The genotype of the second generation offspring of two types of transgenic mice accorded with the Mendelian laws and C/EBPα-/- mice were generated. C/EBPα-/- mice had lower mRNA and protein expressions of C/EBPα in renal tissues and no significant renal morphologic changes compared with WT mice.Conclusion·Cre-loxp system can successfully knock out the C/EBPα gene in mouse renal tubular epithelial cells. C/EBPα-/- mice have no significant renal morphologic changes in baseline condition.

Key words: Cre-loxp system； C/EBPα； conditional knockout, renal tubular epithelial cells