title: Expression of intracellular interferon-alpha confers antiviral properties in transfected bovine fetal fibroblasts and does not affect the full development of SCNT embryos.
abstract: Foot-and-mouth disease, one of the most significant diseases of dairy herds, has substantial effects on farm economics, and currently, disease control measures are limited. In this study, we constructed a vector with a human interferon-α (hIFN-α) (without secretory signal sequence) gene cassette containing the immediate early promoter of human cytomegalovirus. Stably transfected bovine fetal fibroblasts were obtained by G418 selection, and hIFN-α transgenic embryos were produced by somatic cell nuclear transfer (SCNT). Forty-six transgenic embryos were transplanted into surrogate cows, and five cows (10.9%) became pregnant. Two male cloned calves were born. Expression of hIFN-α was detected in transfected bovine fetal fibroblasts, transgenic SCNT embryos, and different tissues from a transgenic SCNT calf at two days old. In transfected bovine fetal fibroblasts, expression of intracellular IFN-α induced resistance to vesicular stomatitis virus infection, increased apoptosis, and induced the expression of double-stranded RNA-activated protein kinase gene (PKR) and the 2'-5'-oligoadenylate synthetase gene (2'-5' OAS), which are IFN-inducible genes with antiviral activity. Analysis by qRT-PCR showed that the mRNA expression levels of PKR, 2'-5' OAS, and P53 were significantly increased in wild-type bovine fetal fibroblasts stimulated with extracellular recombinant human IFN-α-2b, showing that intracellular IFN-α induces biological functions similar to extracellular IFN-α. In conclusion, expression of intracellular hIFN-α conferred antiviral properties in transfected bovine fetal fibroblasts and did not significantly affect the full development of SCNT embryos. Thus, IFN-α transgenic technology may provide a revolutionary way to achieve elite breeding of livestock.
authors.affiliation: Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China; State Key Laboratories of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing, China.
authors.forename: Dawei
authors.initials: D
authors.lastname: Yu
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Science, Changchun, China.
authors.forename: Shoufeng
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Weihua
authors.initials: W
authors.lastname: Du
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Science, Changchun, China.
authors.forename: Jinxia
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Zongxing
authors.initials: Z
authors.lastname: Fan
authors.affiliation: Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Haisheng
authors.initials: H
authors.lastname: Hao
authors.affiliation: Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Xueming
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Tong
authors.initials: T
authors.lastname: Qin
authors.affiliation: Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Huabin
authors.initials: H
authors.lastname: Zhu
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002417:Cattle / Q000235:genetics* / Q000276:immunology; D002418:Cattle Diseases / Q000276:immunology; D002478:Cells, Cultured; D060467:Disease Resistance; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000821:virology; D005536:Foot-and-Mouth Disease / Q000276:immunology; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental / Q000276:immunology; D006801:Humans; D007113:Immunity, Innate; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D053652:Nuclear Transfer Techniques; D011247:Pregnancy; D014162:Transfection; D018116:Vesiculovirus / Q000276:immunology
chemical_list: D016898:Interferon-alpha