title: Direct production of mouse disease models by embryo microinjection of TALENs and oligodeoxynucleotides.
abstract: The study of genetic disease mechanisms relies mostly on targeted mouse mutants that are derived from engineered embryonic stem (ES) cells. Nevertheless, the establishment of mutant ES cells is laborious and time-consuming, restricting the study of the increasing number of human disease mutations discovered by high-throughput genomic analysis. Here, we present an advanced approach for the production of mouse disease models by microinjection of transcription activator-like effector nucleases (TALENs) and synthetic oligodeoxynucleotides into one-cell embryos. Within 2 d of embryo injection, we created and corrected chocolate missense mutations in the small GTPase RAB38; a regulator of intracellular vesicle trafficking and phenotypic model of Hermansky-Pudlak syndrome. Because ES cell cultures and targeting vectors are not required, this technology enables instant germline modifications, making heterozygous mutants available within 18 wk. The key features of direct mutagenesis by TALENs and oligodeoxynucleotides, minimal effort and high speed, catalyze the generation of future in vivo models for the study of human disease mechanisms and interventions.
authors: Institute of Developmental Genetics, Helmholtz Zentrum München, 85764 Munich, Germany.
authors: Benedikt
authors: B
authors: Wefers
authors: Melanie
authors: M
authors: Meyer
authors: Oskar
authors: O
authors: Ortiz
authors: Martin
authors: M
authors: Hrabé de Angelis
authors: Jens
authors: J
authors: Hansen
authors: Wolfgang
authors: W
authors: Wurst
authors: Ralf
authors: R
authors: Kühn
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002970:Cleavage Stage, Ovum; D003851:Deoxyribonucleases / Q000008:administration & dosage / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D030342:Genetic Diseases, Inborn / Q000235:genetics*; D005821:Genetic Techniques; D005822:Genetic Vectors; D018095:Germ-Line Mutation; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008845:Microinjections; D008969:Molecular Sequence Data; D016296:Mutagenesis; D020125:Mutation, Missense; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage* / Q000235:genetics*; D011247:Pregnancy; D012689:Sequence Homology, Nucleic Acid; D020691:rab GTP-Binding Proteins / Q000235:genetics
chemical_list: D009838:Oligodeoxyribonucleotides; D003851:Deoxyribonucleases; C498786:Rab38 protein, mouse; D020691:rab GTP-Binding Proteins