title: Production of transgenic goats expressing human coagulation factor IX in the mammary glands after nuclear transfer using transfected fetal fibroblast cells.
abstract: There are growing numbers of recombinant proteins that have been expressed in milk. Thus one can consider the placement of any gene of interest under the control of the regulatory elements of a milk protein gene in a dairy farm animal. Among the transgene introducing techniques, only nuclear transfer (NT) allows 100 % efficiency and bypasses the mosaicism associated with counterpart techniques. In this study, in an attempt to produce a transgenic goat carrying the human coagulation factor IX (hFIX) transgene, goat fetal fibroblasts were electroporated with a linearized marker-free construct in which the transgene was juxtaposed to β-casein promoter designed to secret the recombinant protein in goat milk. Two different lines of transfected cells were used as donors for NT to enucleated oocytes. Two transgenic goats were liveborn. DNA sequencing of the corresponding transgene locus confirmed authenticity of the cloning procedure and the complementary experiments on the whey demonstrated expression of human factor IX in the milk of transgenic goats. In conclusion, our study has provided the groundwork for a prosperous and promising approach for large-scale production and therapeutic application of hFIX expressed in transgenic goats.
authors.affiliation: Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, 12 Hafez St, Banihashem St, Resalat St, PO Box 19395-4644, Tehran, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Amiri Yekta
authors.forename: Azam
authors.initials: A
authors.lastname: Dalman
authors.forename: Poopak
authors.initials: P
authors.lastname: Eftekhari-Yazdi
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Sanati
authors.forename: Abdol Hossein
authors.initials: AH
authors.lastname: Shahverdi
authors.forename: Rahman
authors.initials: R
authors.lastname: Fakheri
authors.forename: Hamed
authors.initials: H
authors.lastname: Vazirinasab
authors.forename: Mohammad Taghi
authors.initials: MT
authors.lastname: Daneshzadeh
authors.forename: Mahdi
authors.initials: M
authors.lastname: Vojgani
authors.forename: Alireza
authors.initials: A
authors.lastname: Zomorodipour
authors.forename: Nayeralsadat
authors.initials: N
authors.lastname: Fatemi
authors.forename: Zeinab
authors.initials: Z
authors.lastname: Vahabi
authors.forename: Shahab
authors.initials: S
authors.lastname: Mirshahvaladi
authors.forename: Fariba
authors.initials: F
authors.lastname: Ataei
authors.forename: Elmira
authors.initials: E
authors.lastname: Bahraminejad
authors.forename: Najmehsadat
authors.initials: N
authors.lastname: Masoudi
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Rezazadeh Valojerdi
authors.forename: Hamid
authors.initials: H
authors.lastname: Gourabi
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D005164:Factor IX / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006041:Goats / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008321:Mammary Glands, Animal / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000254:growth & development; D014162:Transfection
chemical_list: D005164:Factor IX