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•], targeted mammals were generated by nuclear transfer (the first were reported in [8]). Two more loci were successfully targeted. However, the high targeting frequency reported in [8] was not observed, possibly because the expression level of the target genes did not allow selection conditions as stringent as the ones used in that study.
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•], targeted mammals were generated by nuclear transfer (the first were reported in [8]). Two more loci were successfully targeted. However, the high targeting frequency reported in [8] was not observed, possibly because the expression level of the target genes did not allow selection conditions as stringent as the ones used in that study.
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A GFP marker gene was shown correctly expressed in cloned medaka, and transmitted to offspring at a Mendelian ratio. However, non-cultured embryonic cells were used for cloning, which may not be suitable for targeting manipulation.
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Wakamatsu Y., Ju B., Pristyaznhyuk I., Niwa K., Ladygina T., Kinoshita M., Araki K., Ozato K. Fertile and diploid nuclear transplants derived from embryonic cells of a small laboratory fish, medaka (Oryzias latipes). Proc. Natl. Acad. Sci. U.S.A. 98:2001;1071-1076 A GFP marker gene was shown correctly expressed in cloned medaka, and transmitted to offspring at a Mendelian ratio. However, non-cultured embryonic cells were used for cloning, which may not be suitable for targeting manipulation.
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Cloned fish were generated from primary as well as long-term cultured fibroblasts. Long-term cultured cells would facilitate gene-targeting manipulations.
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Lee K.Y., Huang H., Ju B., Yang Z., Lin S. Cloned zebrafish by nuclear transfer from long-term-cultured cells. Nat. Biotechnol. 20:2002;795-799 Cloned fish were generated from primary as well as long-term cultured fibroblasts. Long-term cultured cells would facilitate gene-targeting manipulations.
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This is the first case of targeted mutagenesis in Drosophila by the whole organism targeting scheme. It also described a novel method that significantly shortened the screening process for targeted events.
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Rong Y.S., Golic K.G. A targeted gene knockout in Drosophila. Genetics. 157:2001;1307-1312 This is the first case of targeted mutagenesis in Drosophila by the whole organism targeting scheme. It also described a novel method that significantly shortened the screening process for targeted events.
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In this report, we firmly established the notion that the whole-organism targeting scheme should be generally applicable for targeted mutagenesis in Drosophila. Five loci were successfully modified by two schemes on the basis of ends-in targeting, more importantly the 'two-step allelic replacement' scheme. In this scheme, a second rare-cutting endonuclease, I-CreI, was expressed to induce recombination between the target gene duplication, which gave rise to a single copy of the target gene carrying a point mutation. Several parameters were also identified that affected gene-targeting efficiency.
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Isolation of Su(var)3-7 mutations by homologous recombination in Drosophila melanogaster
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This is the first report of a targeting success outside of the Golic laboratory. For a gene that has been subjected to ten years of random mutagenesis, a mutant was finally recovered by gene targeting.
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Seum C., Pauli D., Delattre M., Jaquet Y., Spierer A., Spierer P. Isolation of Su(var)3-7 mutations by homologous recombination in Drosophila melanogaster. Genetics. 161:2002;1125-1136 This is the first report of a targeting success outside of the Golic laboratory. For a gene that has been subjected to ten years of random mutagenesis, a mutant was finally recovered by gene targeting.
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