Abstract
Genome editing has revolutionized plant research and plant breeding by enabling precise genome manipulation. In particular, the application of type II CRISPR-Cas9 systems to genome editing has proved an important milestone, accelerating genetic engineering and the analysis of gene function. On the other hand, the potential of other types of CRISPR-Cas systems, especially many of the most abundant type I CRISPR-Cas systems, remains unexplored. We recently developed a novel genome editing tool, TiD, based on the type I-D CRISPR-Cas system. In this chapter, we describe a protocol for genome editing of plant cells using TiD. This protocol allows the application of TiD to induce short insertion and deletions (indels) or long-range deletions at target sites with high specificity in tomato cells.
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Acknowledgments
The work is supported by New Energy and Industrial Technology Development Organization (NEDO) and Japan Science and Technology Agency (JST) grant number JPMJPF2010 (to Y.O.), Adaptable and Seamless Technology transfer Program through Target-driven R&D (A-STEP) (to K.O.), Core Research for Evolutional Science and Technology (CREST) (to K.O.), and JSPS KAKENHI Grant number 22Â K06192 (to N.W.).
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Wada, N., Osakabe, K., Osakabe, Y. (2023). Type I-D CRISPR System-Mediated Genome Editing in Plants. In: Yang, B., Harwood, W., Que, Q. (eds) Plant Genome Engineering. Methods in Molecular Biology, vol 2653. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3131-7_2
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DOI: https://doi.org/10.1007/978-1-0716-3131-7_2
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