title: Myostatin gene mutated mice induced with tale nucleases.
abstract: Myostain gene (MSTN) is expressed primarily in skeletal muscle, and negatively regulates skeletal muscle mass; it has been suggested that mice with MSTN inhibition have reduced adiposity and improved insulin sensitivity. Therefore, it is important to establish a fast and effective gene editing method. In this report, we established the myostatin mutated-mouse model by microinjection of Transcription Activator-Like Effector Nucleases (TALENs) mRNA within the mouse fertilized oocytes and achieved high rates of mutagenesis of the mouse MSTN in C57BL/6J. Six of 45 born mice carried target mutations and we appointed one as the parental mating with wild mouse to produce the F1 and backcross to produce the F2 generation. All the mutations of the mice were examined quickly and efficiently by high-resolution melting curve analysis (HRMA) and then verified by direct sequencing. We obtained the homozygous of the F2 generation which transmitted the mutant alleles to the progeny with 100% efficiency. Mutant mice exhibited increases in muscle mass comparable to those observed in wild-type mice. Therefore, combining TALEN-mediated gene targeting with HRMA technology is a superior method of constructing genetically modified mice through microinjection in the mouse fertilized oocytes with high efficiency and short time of selection.
authors.affiliation: a State Key Laboratory of Genetic Engineering, Fudan-VARI Genetic Epidemiology Center and MOE Key Laboratory of Contemporary Anthropology, School of Life Sciences , Fudan University , Shanghai , China.
authors.forename: Fangfang
authors.initials: F
authors.lastname: Zhou
authors.forename: Ruilin
authors.initials: R
authors.lastname: Sun
authors.forename: Hongyan
authors.initials: H
authors.lastname: Chen
authors.forename: Jian
authors.initials: J
authors.lastname: Fei
authors.forename: Daru
authors.initials: D
authors.lastname: Lu
mesh_terms: D000818:Animals; D001483:Base Sequence; D004252:DNA Mutational Analysis; D003851:Deoxyribonucleases / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000473:pathology*; D009154:Mutation; D055435:Myostatin / Q000235:genetics*; D010641:Phenotype; D014157:Transcription Factors / Q000235:genetics*
chemical_list: C525912:Mstn protein, mouse; D055435:Myostatin; D014157:Transcription Factors; D003851:Deoxyribonucleases
keywords: Gene editing; HRMA; Myostatin; TALEN