PRODUCTION OF NUCLEAR TRANSFER-DERIVED SWINE THAT EXPRESS THE ENHANCED GREEN FLUORESCENT PROTEIN

Abstract

The ability to add or delete specific genes in swine will likely provide considerable benefits not just to agriculture but also to medicine, where pigs have potential as models for human disease and as organ donors. Here we have transferred nuclei from a genetically modified fibroblast cell line to porcine oocytes, matured in vitro under defined culture conditions, to create piglets expressing enhanced green fluorescent protein. The nuclear transfer-derived piglets were of normal size, although some mild symptoms of “large offspring syndrome” were evident. These experiments represent a next step towards creating swine with more useful genetic modifications.

ACKNOWLEDGMENTS

The authors thank A.W.S. Chan for the generous gift of replication defective retroviral vector containing the EGFP, Edmund Rucker for running the microsatellites and the PCR for EGFP on the resulting piglets, and U.S. Pig Genome Coordinator, Max Rothschild for the primers for the microsatellites. The authors would also like to thank Ed Brown and Daniel Liske for care of the surrogate gilts during gestation, Tom Cantley for help with surgical embryo transfers, and Kristin Whitworth, Rami Woods, Zhixin Wei, Jennifer Luth, Lisa Overman, Ryan Cabot, and Dave Wax for helping to care for the piglets after delivery, and Jim Curley for photos in Figure 1a–d. We acknowledge funding from the F.B. Miller fund (Department of Animal Sciences, University of Missouri-Columbia) to H.T.C., funding from the NIH DHHS via R01 RR13428 to R.S.P. and B.N.D., and Food for the 21^st Century. D.B.C. is the recipient of a Pathobiology fellowship via funding from NIH DHHS T32 RR07004.