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Targeted disruption of the porcine immunoglobulin kappa light chain locus

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Abstract

Inactivation of the endogenous pig immunoglobulin (Ig) loci, and replacement with their human counterparts, would produce animals that could alleviate both the supply and specificity issues of therapeutic human polyclonal antibodies (PAbs). Platform genetics are being developed in pigs that have all endogenous Ig loci inactivated and replaced by human counterparts, in order to address this unmet clinical need. This report describes the deletion of the porcine kappa (κ) light chain constant (Cκ) region in pig primary fetal fibroblasts (PPFFs) using gene targeting technology, and the generation of live animals from these cells via somatic cell nuclear transfer (SCNT) cloning. There are only two other targeted loci previously published in swine, and this is the first report of a targeted disruption of an Ig light chain locus in a livestock species. Pigs with one targeted Cκ allele (heterozygous knockout or ±) were bred together to generate Cκ homozygous knockout (−/−) animals. Peripheral blood mononuclear cells (PBMCs) and mesenteric lymph nodes (MLNs) from Cκ −/− pigs were devoid of κ-containing Igs. Furthermore, there was an increase in lambda (λ) light chain expression when compared to that of wild-type littermates (Cκ +/+). Targeted inactivation of the Ig heavy chain locus has also been achieved and work is underway to inactivate the pig lambda light chain locus.

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Acknowledgments

We thank J. MacPherson, B. Gragg, W. Lucero, T. Akers, H. Bishop, and C. Steger for technical contributions to embryo transfer, animal husbandry, and sample procurement; staff at the Virginia Maryland Regional Veterinary College for physical examination of piglets, necropsy report consulting, and histology processing, especially K. Gragg, A. Richardson, and J. Jones for administrative assistance. This work was supported in part by grants from the Defense Advanced Research Projects Agency (DARPA) and the National Institute of Standards and Technology’s (NIST) Advanced Technology Program (ATP) and the National Institutes of Health. We also acknowledge the National Science Foundation (NSF) and National Porkboard for their support in this research.

Competing Interests Statement

J.R., M.M., C.P., T.V., S.B., Y.D., J.M., S.C., A.D., J.Boone, P.J., A.V., I.P., K.W., and D.A. are or were employees of Revivicor, Inc.

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Author notes

  1. K. Wells

    Present address: Division of Animal Sciences, University of Missouri, 920 East Campus Drive, Columbia, MO, 65211, USA

Authors and Affiliations

  1. Revivicor, Inc., 1700 Kraft Drive, Blacksburg, VA, 24060, USA

    J. Ramsoondar, M. Mendicino, C. Phelps, T. Vaught, S. Ball, J. Monahan, S. Chen, A. Dandro, J. Boone, P. Jobst, A. Vance, I. Polejaeva, Y. Dai, D. Ayares & K. Wells

  2. Department of Microbiology and Interdisciplinary Immunology Program, University of Iowa, 3-550 Bowen Science Building, Iowa City, IA, 52242, USA

    N. Wertz & J. Butler

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  1. J. Ramsoondar
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  2. M. Mendicino
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  3. C. Phelps
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  4. T. Vaught
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  5. S. Ball
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  6. J. Monahan
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  7. S. Chen
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  8. A. Dandro
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  9. J. Boone
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  10. P. Jobst
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  11. A. Vance
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  12. N. Wertz
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  13. I. Polejaeva
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  14. J. Butler
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  15. Y. Dai
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  16. D. Ayares
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Corresponding authors

Correspondence to D. Ayares or K. Wells.

Additional information

Ramsoondar and Mendicino contributed equally to this work.

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Ramsoondar, J., Mendicino, M., Phelps, C. et al. Targeted disruption of the porcine immunoglobulin kappa light chain locus. Transgenic Res 20, 643–653 (2011). https://doi.org/10.1007/s11248-010-9445-y

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  • DOI: https://doi.org/10.1007/s11248-010-9445-y

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