title: Transgenic-cloned pigs systemically expressing red fluorescent protein, Kusabira-Orange.
abstract: Genetically engineered pigs with cell markers such as fluorescent proteins are highly useful in lines of research that include the tracking of transplanted cells or tissues. In this study, we produced transgenic-cloned pigs carrying a gene for the newly developed red fluorescent protein, humanized Kusabira-Orange (huKO), which was cloned from the coral stone Fungia concinna. The nuclear transfer embryos, reconstructed with fetal fibroblast cells that had been transduced with huKO cDNA using retroviral vector D Delta Nsap, developed efficiently in vitro into blastocysts (28.0%, 37/132). Nearly all (94.6%, 35/37) of the cloned blastocysts derived from the transduced cells exhibited clear huKO gene expression. A total of 429 nuclear transfer embryos were transferred to four recipients, all of which became pregnant and gave birth to 18 transgenic-cloned offspring in total. All of the pigs highly expressed huKO fluorescence in all of the 23 organs and tissues analyzed, including the brain, eyes, intestinal and reproductive organs, skeletal muscle, bone, skin, and hoof. Furthermore, such expression was also confirmed by histological analyses of various tissues such as pancreatic islets, renal corpuscles, neuronal and glial cells, the retina, chondrocytes, and hematopoietic cells. These data demonstrate that transgenic-cloned pigs exhibiting systemic red fluorescence expression can be efficiently produced by nuclear transfer of somatic cells retrovirally transduced with huKO gene.
authors: Laboratory of Developmental Engineering, Department of Life Science, School of Agriculture, Meiji University, Kawasaki 214-8571, Kanagawa, Japan.
authors: Hitomi
authors: H
authors: Matsunari
authors: Masafumi
authors: M
authors: Onodera
authors: Norihiro
authors: N
authors: Tada
authors: Hideki
authors: H
authors: Mochizuki
authors: Satoshi
authors: S
authors: Karasawa
authors: Erika
authors: E
authors: Haruyama
authors: Naoki
authors: N
authors: Nakayama
authors: Hitoshi
authors: H
authors: Saito
authors: Satoshi
authors: S
authors: Ueno
authors: Mayuko
authors: M
authors: Kurome
authors: Atsushi
authors: A
authors: Miyawaki
authors: Hiroshi
authors: H
authors: Nagashima
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D019976:Cloning, Organism; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006801:Humans; D049408:Luminescent Agents / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000502:physiology; D011247:Pregnancy; D013552:Swine; D014018:Tissue Distribution; D000097573:Red Fluorescent Protein
chemical_list: D049408:Luminescent Agents; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein