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An allelic series for the leptin receptor gene generated by CRE and FLP recombinase

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Abstract

Body weight regulation is mediated through several major signaling pathways, some of which have been delineated by positional cloning of spontaneous genetic mutations in mice. Leprdb/db mice are obese due to a defect in the signaling portion of the leptin receptor, which has led to extensive study of this highly conserved system over the past several years. We have created an allelic series at Lepr for the further examination of LEPR signaling phenotypes using both the FLP/frt and CRE/loxP systems. By inserting a frt-PGK-neo-frt sequence in Lepr intron 16, we have generated a conditional gene repair Lepr allele (Lepr-neo) that elicits morbid obesity, diabetes, and infertility in homozygous mice, recapitulating the obesity syndrome of Leprdb/db mice. Thus, in vivo excision of the PGK-neo cassette with a FLP recombinase transgene restores the lean and fertile phenotype to Leprflox/flox mice. In the same construct, we have also inserted loxP sites that flank Lepr coding exon 17, a region that encodes a JAK docking site required for STAT3 signaling. CRE-mediated excision of Lepr coding exon 17 from Lepr with a frameshift in subsequent exons results in a syndrome of obesity, diabetes, and infertility in LeprΔ17/Δ17 mice, which is indistinguishable from Leprneo/neo and Leprdb/db mice. We conclude that suppression of Lepr gene expression by PGK-neo is phenotypically equivalent to deletion of the Lepr signaling motifs, and therefore the Leprneo/neo mouse may be used to investigate conditional gene repair of Lepr signaling deficiency.

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Acknowledgments

This work was supported by NIH grants DK 57621 (S.C.C.), DK 61229 (J.E.M.), training grant DK 007647, center grants DK 26687 and DK 63608, and NCI grant CA 97403 (T.L.). Sequences for Leprneo, Leprflox and LeprΔ17 have been deposited in GenBank (Accession nos. AY589053, AY589054, and AY589055, respectively).

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Authors and Affiliations

  1. Department of Pediatrics, Division of Molecular Genetics, Columbia University College of Physicians and Surgeons, New York, New York, 10032, USA

    Julie E. McMinn, Shun-Mei Liu, Sandra Eiden & Streamson C. Chua Jr.

  2. Department of Physiology, University of Tennessee, Memphis, Tennessee, 38163, USA

    Ioannis Dragatsis & Paula Dietrich

  3. Department of Anatomy and Cell Biology, Columbia University College of Physicians and Surgeons, New York, New York, 10032, USA

    Thomas Ludwig

  4. Institute of Cancer Genetics, Columbia University College of Physicians and Surgeons, New York, New York, 10032, USA

    Thomas Ludwig

  5. Institute of Human Nutrition, Columbia University College of Physicians and Surgeons, New York, New York, 10032, USA

    Streamson C. Chua Jr.

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  1. Julie E. McMinn
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  2. Shun-Mei Liu
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  3. Ioannis Dragatsis
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  4. Paula Dietrich
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  5. Thomas Ludwig
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  7. Streamson C. Chua Jr.
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Correspondence to Streamson C. Chua Jr..

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McMinn, J.E., Liu, SM., Dragatsis, I. et al. An allelic series for the leptin receptor gene generated by CRE and FLP recombinase. Mamm Genome 15, 677–685 (2004). https://doi.org/10.1007/s00335-004-2340-1

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  • DOI: https://doi.org/10.1007/s00335-004-2340-1

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