title: Development of transgenic minipigs with expression of antimorphic human cryptochrome 1.
abstract: Minipigs have become important biomedical models for human ailments due to similarities in organ anatomy, physiology, and circadian rhythms relative to humans. The homeostasis of circadian rhythms in both central and peripheral tissues is pivotal for numerous biological processes. Hence, biological rhythm disorders may contribute to the onset of cancers and metabolic disorders including obesity and type II diabetes, amongst others. A tight regulation of circadian clock effectors ensures a rhythmic expression profile of output genes which, depending on cell type, constitute about 3-20% of the transcribed mammalian genome. Central to this system is the negative regulator protein Cryptochrome 1 (CRY1) of which the dysfunction or absence has been linked to the pathogenesis of rhythm disorders. In this study, we generated transgenic Bama-minipigs featuring expression of the Cys414-Ala antimorphic human Cryptochrome 1 mutant (hCRY1(AP)). Using transgenic donor fibroblasts as nuclear donors, the method of handmade cloning (HMC) was used to produce reconstructed embryos, subsequently transferred to surrogate sows. A total of 23 viable piglets were delivered. All were transgenic and seemingly healthy. However, two pigs with high transgene expression succumbed during the first two months. Molecular analyzes in epidermal fibroblasts demonstrated disturbances to the expression profile of core circadian clock genes and elevated expression of the proinflammatory cytokines IL-6 and TNF-α, known to be risk factors in cancer and metabolic disorders.
authors.affiliation: BGI-Shenzhen, Shenzhen, Guangdong, China ; ShenZhen Engineering Laboratory for Genomics-Assisted Animal Breeding, BGI-Shenzhen, Shenzhen, Guangdong, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Liu
authors.forename: Yong
authors.initials: Y
authors.lastname: Li
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wei
authors.forename: Chunxin
authors.initials: C
authors.lastname: Liu
authors.forename: Lars
authors.initials: L
authors.lastname: Bolund
authors.forename: Gábor
authors.initials: G
authors.lastname: Vajta
authors.forename: Hongwei
authors.initials: H
authors.lastname: Dou
authors.forename: Wenxian
authors.initials: W
authors.lastname: Yang
authors.forename: Ying
authors.initials: Y
authors.lastname: Xu
authors.forename: Jing
authors.initials: J
authors.lastname: Luan
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Huanming
authors.initials: H
authors.lastname: Yang
authors.forename: Nicklas Heine
authors.initials: NH
authors.lastname: Staunstrup
authors.forename: Yutao
authors.initials: Y
authors.lastname: Du
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology / Q000235:genetics* / Q000254:growth & development; D057906:Circadian Clocks / Q000235:genetics*; D002940:Circadian Rhythm / Q000235:genetics*; D019976:Cloning, Organism; D056931:Cryptochromes / Q000235:genetics* / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010507:Periodicity; D013552:Swine; D013556:Swine, Miniature / Q000196:embryology / Q000235:genetics* / Q000254:growth & development; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: C538963:CRY1 protein, human; D056931:Cryptochromes; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha