Reference - Detail
| RRC ID | 33656 |
|---|---|
| Author | Kimura Y, Hisano Y, Kawahara A, Higashijima S. |
| Title | Efficient generation of knock-in transgenic zebrafish carrying reporter/driver genes by CRISPR/Cas9-mediated genome engineering. |
| Journal | Sci Rep |
| Abstract |
The type II bacterial CRISPR/Cas9 system is rapidly becoming popular for genome-engineering due to its simplicity, flexibility, and high efficiency. Recently, targeted knock-in of a long DNA fragment via homology-independent DNA repair has been achieved in zebrafish using CRISPR/Cas9 system. This raised the possibility that knock-in transgenic zebrafish could be efficiently generated using CRISPR/Cas9. However, how widely this method can be applied for the targeting integration of foreign genes into endogenous genomic loci is unclear. Here, we report efficient generation of knock-in transgenic zebrafish that have cell-type specific Gal4 or reporter gene expression. A donor plasmid containing a heat-shock promoter was co-injected with a short guide RNA (sgRNA) targeted for genome digestion, a sgRNA targeted for donor plasmid digestion, and Cas9 mRNA. We have succeeded in establishing stable knock-in transgenic fish with several different constructs for 4 genetic loci at a frequency being exceeding 25%. Due to its simplicity, design flexibility, and high efficiency, we propose that CRISPR/Cas9-mediated knock-in will become a standard method for the generation transgenic zebrafish. |
| Volume | 4 |
| Pages | 6545 |
| Published | 2014-10-8 |
| DOI | 10.1038/srep06545 |
| PII | srep06545 |
| PMID | 25293390 |
| PMC | PMC4189020 |
| MeSH | Animals Animals, Genetically Modified CRISPR-Cas Systems* Gene Knock-In Techniques* Gene Targeting Genetic Engineering* Genome Organisms, Genetically Modified / genetics* RNA, Guide, CRISPR-Cas Systems / genetics Zebrafish / genetics* |
| IF | 3.998 |
| Times Cited | 136 |
| WOS Category | MULTIDISCIPLINARY SCIENCES |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 28 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Zebrafish | Tg[vglut2a:GFP] UAS:RFP |