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Disruption of the Survival Motor Neuron (SMN) gene in pigs using ssDNA

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Abstract

Spinal Muscular Atrophy (SMA) is an autosomal recessive neurodegenerative disease that is a result of a deletion or mutation of the SMN1 (Survival Motor Neuron) gene. A duplicated and nearly identical copy, SMN2, serves as a disease modifier as increasing SMN2 copy number decreases the severity of the disease. Currently many therapeutic approaches for SMA are being developed. Therapeutic strategies aim to modulate splicing of SMN2-derived transcripts, increase SMN2 gene expression, increase neuro-protection of motor neurons, stabilize the SMN protein, replace the SMN1 gene and reconstitute the motor neuron population. It is our goal to develop a pig animal model of SMA for the development and testing of therapeutics and evaluation of toxicology. In the development of a SMA pig model, it was important to demonstrate that the human SMN2 gene would splice appropriately as the model would be based on the presence of the human SMN2 transgene. In this manuscript, we show splicing of the human SMN1 and SMN2 mini-genes in porcine cells is consistent with splicing in human cells, and we report the first genetic knockout of a gene responsible for a neurodegenerative disease in a large animal model using gene targeting with single-stranded DNA and somatic cell nuclear transfer.

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Acknowledgments

Thank you to members of the C. Lorson, K. Wells and R. Prather laboratories for their assistance and generosity especially Martha Bennett, Chad O’Gorman, Erik Osman, Armedia Stump, Hans Rindt, August Rieke and Jason Dowell. This work was supported by FightSMA and National Institutes of Health [grant number NS059510 to MAL].

Conflict of interest

The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Department of Veterinary Pathobiology, Bond Life Sciences Center, University of Missouri, 1201 Rollins Road, Room 440C, Columbia, MO, 65211-7310, USA

    Monique A. Lorson & Christian L. Lorson

  2. Division of Animal Science, Animal Science Research Center, University of Missouri, 920 East Campus Drive, Columbia, MO, 65211, USA

    Lee D. Spate, Melissa S. Samuel & Clifton N. Murphy

  3. Division of Animal Science, Animal Science Research Center, University of Missouri, Columbia, MO, 65211, USA

    Randall S. Prather & Kevin D. Wells

  4. Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, MO, 65211, USA

    Christian L. Lorson

  5. Department of Veterinary Pathobiology, Bond Life Sciences Center, University of Missouri, 1201 Rollins Road, Columbia, MO, 65211-7310, USA

    Christian L. Lorson

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  1. Monique A. Lorson
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  2. Lee D. Spate
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  3. Melissa S. Samuel
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  4. Clifton N. Murphy
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  5. Christian L. Lorson
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  6. Randall S. Prather
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  7. Kevin D. Wells
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Correspondence to Monique A. Lorson.

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Lorson, M.A., Spate, L.D., Samuel, M.S. et al. Disruption of the Survival Motor Neuron (SMN) gene in pigs using ssDNA. Transgenic Res 20, 1293–1304 (2011). https://doi.org/10.1007/s11248-011-9496-8

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  • DOI: https://doi.org/10.1007/s11248-011-9496-8

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