NDLI: GRK5-Knockout Mice Generated by TALEN-Mediated Gene Targeting.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Nanjidsuren, Tsevelmaa Park, Chae-Won Sim, Bo-Woong Kim, Sun-Uk Chang, Kyu-Tae Kang, Myung-Hwa Min, Kwan-Sik
Description	Author Affiliation: Nanjidsuren T ( a Animal Biotechnology, Graduate School of Future Convergence Technology , Institute of Genetic Engineering, Hankyong National University , Anseong , Republic of Korea.); Park CW ( a Animal Biotechnology, Graduate School of Future Convergence Technology , Institute of Genetic Engineering, Hankyong National University , Anseong , Republic of Korea.); Sim BW ( b National Research Center , Korea Research Institute of Bioscience and Biotechnology , Ochang , Republic of Korea.); Kim SU ( b National Research Center , Korea Research Institute of Bioscience and Biotechnology , Ochang , Republic of Korea.); Chang KT ( b National Research Center , Korea Research Institute of Bioscience and Biotechnology , Ochang , Republic of Korea.); Kang MH ( c Department of Food and Nutrition , Hoseo University , Asan , Republic of Korea.); Min KS ( a Animal Biotechnology, Graduate School of Future Convergence Technology , Institute of Genetic Engineering, Hankyong National University , Anseong , Republic of Korea.)
Abstract	Transcription activator-like effector nucleases (TALENs) are a new type of engineered nuclease that is very effective for directed gene disruption in any genome sequence. We investigated the generation of mice with genetic knockout (KO) of the G protein-coupled receptor kinase (GRK) 5 gene by microinjection of TALEN mRNA. TALEN vectors were designed to target exons 1, 3, and 5 of the mouse GRK5 gene. Flow cytometry showed that the activity of the TALEN mRNAs targeted to exons 1, 3, and 5 was 8.7%, 9.7%, and 12.7%, respectively. The TALEN mRNA for exon 5 was injected into the cytoplasm of 180 one-cell embryos. Of the 53 newborns, three (5.6%) were mutant founders (F0) with mutations. Two clones from F028 showed a 45-bp deletion and F039 showed the same biallelic non-frame-shifting 3-bp deletions. Three clones from F041 were shown to possess a combination of frame-shifting 2-bp deletions. All of the mutations were transmitted through the germline but not to all progenies (37.5%, 37.5%, and 57.1% for the F028, F039, and F041 lines, respectively). The homozygote GRK5-KO mice for 28 and 41 lines created on F3 progenies and the homozygous genotype was confirmed by PCR, T7E1 assay and sequencing.
File Format	HTM / HTML
ISSN	10495398
Issue Number	4
Journal	Animal Biotechnology
Volume Number	27
e-ISSN	15322378
Language	English
Publisher	Taylor & Francis
Publisher Date	2016-10-01
Publisher Place	Great Britain (UK)
Access Restriction	Subscribed
Subject Keyword	Discipline Zoology Discipline Biotechnology
Content Type	Text
Resource Type	Article
Subject	Animal Science and Zoology Bioengineering Biotechnology

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