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Direct production of mouse disease models by embryo microinjection of TALENs and oligodeoxynucleotides.
Proc. Natl. Acad. Sci. U.S.A. 110, 3782-3787 (2013)
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.
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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
Gene Targeting ; Mouse Genetics ; Knockout ; Knockin; Zinc-finger Nucleases ; Genome ; Mutations ; Knockout ; Rab38 ; Mice ; Discovery ; Effectors
ISSN (print) / ISBN
0027-8424
e-ISSN
1091-6490
Quellenangaben
Band: 110,
Heft: 10,
Seiten: 3782-3787
Verlag
National Academy of Sciences
Nichtpatentliteratur
Publikationen
Begutachtungsstatus
Peer reviewed