Abstract
Zinc finger nucleases (ZFNs) are proteins engineered to make site-specific double-strand breaks (DSBs) in a DNA sequence of interest. Imprecise repair of the ZFN-induced DSBs by the nonhomologous end-joining (NHEJ) pathway results in a spectrum of mutations, such as nucleotide substitutions, insertions, and deletions. Here we describe a method for targeted mutagenesis in Arabidopsis with ZFNs, which are engineered by context-dependent assembly (CoDA). This ZFN-induced mutagenesis method is an alternative to other currently available gene knockout or knockdown technologies and is useful for reverse genetic studies.
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Acknowledgments
This work is supported by grants from the National Science Foundation to D.F.V. (DBI 0923827 and MCB 0209818).
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Qi, Y., Starker, C.G., Zhang, F., Baltes, N.J., Voytas, D.F. (2014). Tailor-Made Mutations in Arabidopsis Using Zinc Finger Nucleases. In: Sanchez-Serrano, J., Salinas, J. (eds) Arabidopsis Protocols. Methods in Molecular Biology, vol 1062. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-580-4_10
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DOI: https://doi.org/10.1007/978-1-62703-580-4_10
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