SURF1 knockout cloned pigs: Early onset of a severe lethal phenotype

Autor: C. Corona, Emanuela Bottani, P. Crociara, I. Di Meo, Mark A. Johnson, Carlo Viscomi, Valeria Tiranti, S. Grifoni, Cesare Galli, Erika Fernandez-Vizarra, C. Casalone, Corinne Quadalti, Giovanna Lazzari, Irina Lagutina, Dario Brunetti, Massimo Zeviani, Raffaele Cerutti, Andrea Perota, Roberto Duchi, Alan J. Robinson
Rok vydání: 2017
Předmět:
0301 basic medicine
Central Nervous System
Male
Nuclear Transfer Techniques
Sus scrofa
Mitochondrion
Animals
Genetically Modified
SURF1 KO
Gene Knockout Techniques
0302 clinical medicine
Genome editing
Leigh syndrome
Mitochondrial disease
Pig
Animals
Animals
Newborn
Behavior
Animal
CRISPR-Cas Systems
Cells
Cultured
Down-Regulation
Electron Transport Complex IV
Female
Fibroblasts
Gene Editing
Humans
Jejunum
Leigh Disease
Membrane Proteins
Mitochondria
Mitochondrial Proteins
Muscle
Skeletal
Primary Cell Culture
Disease Models
Animal
SURF1
Cultured
Skeletal
Phenotype
Muscle
Molecular Medicine
Cells
Genetically Modified
Biology
Article
Andrology
03 medical and health sciences
medicine
Leigh disease
Molecular Biology
Gene
Behavior
Animal
Wild type
Newborn
medicine.disease
030104 developmental biology
MRNA Sequencing
Disease Models
030217 neurology & neurosurgery
Zdroj: Biochimica et Biophysica Acta
ISSN: 0925-4439
Popis: Leigh syndrome (LS) associated with cytochrome c oxidase (COX) deficiency is an early onset, fatal mitochondrial encephalopathy, leading to multiple neurological failure and eventually death, usually in the first decade of life. Mutations in SURF1, a nuclear gene encoding a mitochondrial protein involved in COX assembly, are among the most common causes of LS. LSSURF1 patients display severe, isolated COX deficiency in all tissues, including cultured fibroblasts and skeletal muscle. Recombinant, constitutive SURF1−/− mice show diffuse COX deficiency, but fail to recapitulate the severity of the human clinical phenotype. Pigs are an attractive alternative model for human diseases, because of their size, as well as metabolic, physiological and genetic similarity to humans. Here, we determined the complete sequence of the swine SURF1 gene, disrupted it in pig primary fibroblast cell lines using both TALENs and CRISPR/Cas9 genome editing systems, before finally generating SURF1−/− and SURF1−/+ pigs by Somatic Cell Nuclear Transfer (SCNT). SURF1−/− pigs were characterized by failure to thrive, muscle weakness and highly reduced life span with elevated perinatal mortality, compared to heterozygous SURF1−/+ and wild type littermates. Surprisingly, no obvious COX deficiency was detected in SURF1−/− tissues, although histochemical analysis revealed the presence of COX deficiency in jejunum villi and total mRNA sequencing (RNAseq) showed that several COX subunit-encoding genes were significantly down-regulated in SURF1−/− skeletal muscles. In addition, neuropathological findings, indicated a delay in central nervous system development of newborn SURF1−/− piglets. Our results suggest a broader role of sSURF1 in mitochondrial bioenergetics.
Highlights • The full sequence of pig SURF1 gene was determined. • SURF1 gene was disrupted in pig by gene editing and somatic cell nuclear transfer. • SURF1−/− piglets showed an early onset lethal phenotype. • Mitochondrial bioenergetics was impaired in the skeletal muscle of SURF1−/− pigs.
Databáze: OpenAIRE
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