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Fidelity of a BAC-EGFP transgene in reporting dynamic expression of IL-7Rα in T cells

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Abstract

Interleukin-7 receptor α chain (IL-7Rα)-derived signals are critical for normal T cell development, mature T cell homeostasis, and longevity of memory T cells. IL-7Rα expression in T cells is dynamically regulated at different developmental and antigen-responding stages. However, the molecular mechanism underlying the dynamic regulation is not completely understood. Here we describe generation of a bacterial artificial chromosome (BAC)-based reporter transgenic mouse strain, which contains 210 kb DNA sequence flanking the Il7r locus. We used in vitro validated EGFP reporter and insulator sequences to facilitate the reporter transgene expression. Consistent with endogenous IL-7Rα expression, the BAC transgene was expressed in mature T cells, a portion of natural killer cells but not in mature B cells. In the thymus, the EGFP reporter and endogenous IL-7Rα showed synchronized silencing in CD4+CD8+ double positive stage, were both upregulated in CD4+ or CD8+ single positive thymocytes, and both continued to be co-expressed in naïve T cells in the periphery. Upon encountering antigen, the antigen-specific effector CD8+ T cells downregulated both endogenous IL-7Rα and the EGFP reporter, which were upregulated in synchrony in antigen-specific memory CD8 T cells. These results indicate that the BAC-EGFP transgene reports endogenous IL-7Rα regulation with high fidelity, and further suggest that the 210 kb sequence flanking the Il7r locus contains sufficient genetic information to regulate its expression changes in T lineage cells. Our approach thus represents a critical initial step towards systematic dissection of the cis regulatory elements controlling dynamic IL-7Rα regulation during T cell development and cellular immune responses.

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Acknowledgments

We thank Drs. Donald Court and Neal Copeland (NCI) for providing all the materials for recombineering, Dr. Gary Felsenfeld (NIDDK, NIH) for providing the chicken β-globin insulator. We are grateful to Drs John Harty and Vladimir Badovinac (University of Iowa) for useful discussions and providing LM-Ova and MHC-I-SIINFEKL tetramer. This study is supported by an NIH grant AI042767 to H.H.X.

Conflict of interest

The authors declare no competing financial interests.

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Author notes

  1. Dahai Yu

    Present address: Glaucoma Cell Biology Laboratory of the Iowa Glaucoma Center, Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA, 52242, USA

Authors and Affiliations

  1. Department of Microbiology, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA

    Shuyang Yu, Xinyuan Zhou, Dahai Yu & Hai-Hui Xue

  2. Department of Molecular Physiology and Biophysics, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA

    Jordy J. Hsiao

  3. Department of Human Genetics, China Medical University, 110001, Shenyang, China

    Dahai Yu

  4. Department of Internal Medicine, Division of Molecular Medicine and Genetics, University of Michigan, Ann Arbor, MI, 48109, USA

    Thomas L. Saunders

  5. Interdisciplinary Immunology Graduate Program, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA

    Hai-Hui Xue

  6. 51 Newton Rd. BSB 3-710, Iowa City, IA, 52246, USA

    Hai-Hui Xue

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Correspondence to Hai-Hui Xue.

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Yu, S., Zhou, X., Hsiao, J.J. et al. Fidelity of a BAC-EGFP transgene in reporting dynamic expression of IL-7Rα in T cells. Transgenic Res 21, 201–215 (2012). https://doi.org/10.1007/s11248-011-9508-8

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