| First Author | Nanjidsuren T | Year | 2016 |
| Journal | Anim Biotechnol | Volume | 27 |
| Issue | 4 | Pages | 223-30 |
| PubMed ID | 27565865 | Mgi Jnum | J:278770 |
| Mgi Id | MGI:6356277 | Doi | 10.1080/10495398.2016.1176032 |
| Citation | Nanjidsuren T, et al. (2016) GRK5-Knockout Mice Generated by TALEN-Mediated Gene Targeting. Anim Biotechnol 27(4):223-30 |
| abstractText | 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. |
