Production of α-1,3-galactosyltransferase null pigs by means of nuclear transfer with fibroblasts bearing loss of heterozygosity mutations
| Autor: | Robert J. Hawley, Clifton N. Murphy, Liangxue Lai, August Rieke, Scott Arn, David B. Carter, Jeffery M. Betthauser, Yanhong Hao, David J. J. Ryan, Randall S. Prather, Zhonghua Liu, Gi-Sun Im, David H. Sachs, Donna Kolber-Simonds, Maria Denaro, Erik Forsberg, Monica L. Augenstein, Julia L. Greenstein, Kwang-Wook Park, Greg D. Mell, Steven R. Watt |
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| Rok vydání: | 2004 |
| Předmět: |
Nuclear Transfer Techniques
Multidisciplinary Swine Loss of Heterozygosity Miniature swine Locus (genetics) Fibroblasts Biological Sciences Biology Flow Cytometry Galactosyltransferases Null allele Molecular biology Epitope Cell Line Loss of heterozygosity Blotting Southern Phenotype Inbred strain Null cell Animals Allele |
| Zdroj: | Proceedings of the National Academy of Sciences. 101:7335-7340 |
| ISSN: | 1091-6490 0027-8424 |
| Popis: | Hyperacute rejection of porcine organs by old world primate recipients is mediated through preformed antibodies against galactosyl-α-1,3-galactose (Galα-1,3-Gal) epitopes expressed on the pig cell surface. Previously, we generated inbred miniature swine with a null allele of the α-1,3-galactosyltransferase locus ( GGTA1 ) by nuclear transfer (NT) with gene-targeted fibroblasts. To expedite the generation of GGTA1 null pigs, we selected spontaneous null mutant cells from fibroblast cultures of heterozygous animals for use in another round of NT. An unexpectedly high rate of spontaneous loss of GGTA1 function was observed, with the vast majority of null cells resulting from loss of the WT allele. Healthy piglets, hemizygous and homozygous for the gene-targeted allele, were produced by NT by using fibroblasts that had undergone deletional and crossover/gene conversion events, respectively. Aside from loss of Galα-1,3-Gal epitopes, there were no obvious phenotypic differences between these null piglets and WT piglets from the same inbred lines. In fact, congenital abnormalities observed in the heterozygous NT animals did not reappear in the serially produced null animals. |
| Databáze: | OpenAIRE |
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