Targeted gene knockout by direct delivery of zinc-finger nuclease proteins
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چکیده
منابع مشابه
targeted gene knockout by direct delivery of zinc - finger nuclease proteins
nature methods | ADVANCE ONLINE PUBLICATION | purify in quantities sufficient for analysis in cell culture (data not shown). Following these results, and based on the observation that ZFP DNA-binding domains carry a net positive charge (Fig. 1a), we hypothesized that ZFNs might penetrate the cell in the absence of additional modification. We expressed in Escherichia coli ZFNs designed to target...
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Gene knockout is the most powerful tool for determining gene function or permanently modifying the phenotypic characteristics of a cell. Existing methods for gene disruption are limited by their efficiency, time to completion, and/or the potential for confounding off-target effects. Here, we demonstrate a rapid single-step approach to targeted gene knockout in mammalian cells, using engineered ...
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Zinc-finger nucleases can be designed to cleave arbitrary genomic sequences, which stimulates localized mutagenesis and targeted gene replacement. Both outcomes depend on normal cellular processes of DNA repair, and this study examines the pathways responsible. Gene replacement by homologous recombination depends on the activity of the Rad51 protein. Much of local mutagenesis depends on DNA lig...
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ژورنال
عنوان ژورنال: Nature Methods
سال: 2012
ISSN: 1548-7091,1548-7105
DOI: 10.1038/nmeth.2030