Abstract
Sequence-specific nucleases (SSNs), such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9) system, are powerful tools for understanding gene function and for developing novel traits in plants. In plant species for which transformation and regeneration systems using protoplasts are not yet established, direct delivery to nuclei of SSNs either in the form of RNA or protein is difficult. Thus, Agrobacterium-mediated transformation of SSN expression constructs in cultured cells is a practical means of delivering targeted mutagenesis in some plant species including rice. Because targeted mutagenesis occurs stochastically in transgenic cells and SSN-mediated targeted mutagenesis often leads to no selectable phenotype, identification of highly mutated cell lines is a critical step in obtaining regenerated plants with desired mutations.
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Acknowledgments
We thank M. Mikami and K. Abe for technical assistance. This research was supported by a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan (Genomics for Agricultural Innovation PGE1001), the Council for Science, Technology and Innovation (CSTI), Cross-ministerial Strategic Innovation Promotion Program (SIP), and the NIAS Strategic Research Fund.
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Endo, M., Nishizawa-Yokoi, A., Toki, S. (2016). Targeted Mutagenesis in Rice Using TALENs and the CRISPR/Cas9 System. In: Murata, M. (eds) Chromosome and Genomic Engineering in Plants. Methods in Molecular Biology, vol 1469. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4931-1_9
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DOI: https://doi.org/10.1007/978-1-4939-4931-1_9
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