Homologous Recombination in Zebrafish ES Cells
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Targeted insertion of a plasmid by homologous recombination was demonstrated in zebrafish ES cell cultures. Two selection strategies were used to isolate ES cell colonies that contained targeted plasmid insertions in either the no tail or myostatin I gene. One selection strategy involved the manual isolation of targeted cell colonies that were identified by the loss of fluorescent protein gene expression. A second strategy used the diphtheria toxin A-chain gene in a positive-negative selection approach. Homologous recombination was confirmed by PCR, sequence and Southern blot analysis and colonies isolated using both selection methods were expanded and maintained for multiple passages. The results demonstrate that zebrafish ES cells have potential for use in a cell-mediated gene targeting approach.
Keywordsembryonic stem cells homologous recombination gene targeting zebrafish
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- Biga, PR, Roberts, SB, Iliev, DB, McCauley, LAR, Moon, JS, Collodi, P, Goetz, FW 2005The isolation, characterization, and expression of a novel Gdf11 gene and a second myostatin form in zebrafishDanio rerio Comp Biochem Physiol B Biochem Mol Biol141218230Google Scholar
- Fan, L, Alestrom, A, Alestrom, P, Collodi, P 2004aDevelopment of cell cultures with competency for contributing to the zebrafish germ lineCrit Rev Euk Gene Exp144351Google Scholar
- Fan, L, Crodian, J, Collodi, P 2004cCulture of embryonic stem cell lines from zebrafishMeth Cell Biol76151160Google Scholar
- Hong, Y, Chen, S, Gui, J, Schartl, M 2004Retention of the developmental pluripotency in medaka embryonic stem cells after gene transfer and long-term drug selection for gene targeting in fishTrans Res134150Google Scholar
- McCarrick, JW, Parnes, JR, Seong, RH, Solter, D, Knowles, BB 1993Positive-negative selection gene targeting with the diphtheria toxin A-chain gene in mouse embryonic stem cellsTrans Res2183190Google Scholar