論文 - 詳細
| RRC ID | 30924 |
|---|---|
| 著者 | Guan G, Yan Y, Chen T, Yi M, Ni H, Naruse K, Nagahama Y, Hong Y. |
| タイトル | Nanos3 gene targeting in medaka ES cells. |
| ジャーナル | Int J Biol Sci |
| Abstract |
Gene targeting (GT) by homologous recombination offers the best precision for genome editing in mice. nanos3 is a highly conserved gene and encodes a zinc-finger RNA binding protein essential for germ stem cell maintenance in Drosophila, zebrafish and mouse. Here we report nanos3 GT in embryonic stem (ES) cells of the fish medaka as a lower vertebrate model organism. A vector was designed for GT via homologous recombination on the basis of positive-negative selection (PNS). The ES cell line MES1 after gene transfer and PNS produced 56 colonies that were expanded into ES cell sublines. Nine sublines were GT-positive by PCR genotyping, 4 of which were homologous recombinants as revealed by Southern blot. We show that one of the 4, A15, contains a precisely targeted nanos3 allele without any random events, demonstrating the GT feasibility in medaka ES cells. Importantly, A15 retained all features of undifferentiated ES cells, including stable self-renewal, an undifferentiated phenotype, pluripotency gene expression and differentiation during chimeric embryogenesis. These results provide first evidence that the GT procedure and genuine GT on a chromosomal locus such as nanos3 do not compromise pluripotency in ES cells of a lower vertebrate. |
| 巻・号 | 9(5) |
| ページ | 444-54 |
| 公開日 | 2013-1-1 |
| DOI | 10.7150/ijbs.6507 |
| PII | ijbsv09p0444 |
| PMID | 23678294 |
| PMC | PMC3654493 |
| MeSH | Animals Blotting, Southern Embryonic Stem Cells / metabolism* Gene Targeting / methods* Genetic Engineering / methods* Genetic Vectors / genetics Genotype Mice Oryzias / genetics* Plasmids / genetics Pluripotent Stem Cells / metabolism* RNA-Binding Proteins / genetics* Reverse Transcriptase Polymerase Chain Reaction Sequence Analysis, DNA Species Specificity |
| IF | 4.858 |
| 引用数 | 4 |
| WOS 分野 | BIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| メダカ | database |