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• W2596545265 endingPage "239" @default.
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• W2596545265 abstract "Transgenic pigs are important model organisms in the field of biomedical research, serving as a potential source of appropriately-sized transplantable human organs that carry desirable genetic traits, and as realistic animal models for cardiovascular, respiratory, and reproductive physiology and disease in humans. Cloning by somatic cell nuclear transfer (SCNT) of genetically altered cells is currently the only viable method for producing knock-out and knock-in pigs, since there are no swine embryonic stem cells available. Our lab recently reported on the successful cloning of endothelial nitric oxide synthase (eNOS) overexpressing piglets to better understand the role of this enzyme and nitric oxide (NO) in cardiovascular regulation. A single male clone carrying the eNOS transgene driven by a Tie-2 promoter was bred to eight wild-type sows, yielding a total of 52 piglets, 22 females and 30 males. Three of the offspring (one female and two males from separate litters) died 2 days post-partum. Average litter size was 6.5 ± 1.4 piglets. Inheritance of the eNOS transgene in the piglets was determined by real-time PCR genotyping and immunohistochemical (IHC) analysis of a transgene specific marker. For PCR genotyping, genomic DNA was extracted from tail snips and eNOS-specific real-time PCR primers were used to determine fold-difference in genomic copy number in each pig based on a wild-type eNOS gene standard curve. A total of 40 piglets (78%) with eNOS gene copy greater than 1.35-fold above wild-type were designated as transgenic, with equal percentages of males and females carrying the transgene. The eNOS transgene cassette contains a polyhistidine sequence (6× his) allowing for identification by IHC in tissue sections by using an anti-polyhistidine antibody. The transgene was confirmed to be present in 28 of the piglets (57%) with another 9 piglets having borderline positive staining (18%). Equal percentages of males and females were determined by IHC to carry the transgene. At total of 12 piglets (24%) had conflicting genotypes when PCR and IHC results were compared. Regardless of method used to assess transgene inheritance, a high percentage of offspring were categorized as carrying the eNOS overexpressing sequence. Experiments are underway to determine whether multiple copies of the transgene are present in the genome or whether over-expression of eNOS in the transgenic boar provide sperm with an advantage in fertilization. It has previously been shown that NO (at endogenous concentrations) appears to be necessary for adequate sperm motility, and therefore eNOS over-expression may enhance the migration rate of porcine sperm. The eNOS over-expressing offspring are currently being bred to expand a colony of transgenic miniature pigs that are homozygous for this trait. These pigs will be used in exercise and diet studies to determine how this phenotype is involved in regulating muscle metabolism and cardiovascular function. (poster)" @default.
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• W2596545265 date "2007-07-01" @default.
• W2596545265 modified "2023-09-27" @default.
• W2596545265 title "INHERITANCE PATTERN OF PORCINE ENDOTHELIAL NITRIC OXIDE SYNTHASE (ENOS) OVER-EXPRESSING TRANSGENE IN OFFSPRING OF A CLONED TRANSGENIC BOAR BRED TO WILD-TYPE SOWS" @default.
• W2596545265 doi "https://doi.org/10.1093/biolreprod/77.s1.239a" @default.
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