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• W2146357346 abstract "Gene targeting can be achieved with lentiviral vectors delivering donor sequences along with a nuclease that creates a locus-specific double-strand break (DSB). Therapeutic applications of this system would require an appropriate control of the amount of endonuclease delivered to the target cells, and potentially toxic sustained expression must be avoided. Here, we show that the nuclease can be transferred into cells as a protein associated with a lentiviral vector particle. I-SceI, a prototypic meganuclease from yeast, was incorporated into the virions as a fusion with Vpr, an HIV accessory protein. Integration-deficient lentiviral vectors containing the donor sequences and the I-SceI fusion protein were tested in reporter cells in which targeting events were scored by the repair of a puromycin resistance gene. Molecular analysis of the targeted locus indicated a 2-fold higher frequency of the expected recombination event when the nuclease was delivered as a protein rather than encoded by a separate vector. In both systems, a proportion of clones displayed multiple integrated copies of the donor sequences, either as tandems at the targeted locus or at unrelated loci. These integration patterns were dependent upon the mode of meganuclease delivery, suggesting distinct recombination processes." @default.
• W2146357346 created "2016-06-24" @default.
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• W2146357346 date "2011-06-28" @default.
• W2146357346 modified "2023-09-30" @default.
• W2146357346 title "Efficient gene targeting mediated by a lentiviral vector-associated meganuclease" @default.
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• W2146357346 doi "https://doi.org/10.1093/nar/gkr524" @default.
• W2146357346 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3177226" @default.
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• W2146357346 hasPublicationYear "2011" @default.
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