Optimizing genome editing efficiency in wheat: Effects of heat treatments and different promoters for single guide RNA expression
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- Kishi-Kaboshi Mitsuko
- Institute of Crop Science, National Agriculture and Food Research Organization (NARO)
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- Abe Fumitaka
- Institute of Crop Science, National Agriculture and Food Research Organization (NARO)
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- Kamiya Yoko
- Kihara Institute for Biological Research, Yokohama City University
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- Kawaura Kanako
- Kihara Institute for Biological Research, Yokohama City University
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- Hisano Hiroshi
- Institute of Plant Science and Resources, Okayama University
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- Sato Kazuhiro
- Institute of Plant Science and Resources, Okayama University
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Abstract
<p>Genome editing is a promising method for simultaneously mutagenizing homoeologs in the three subgenomes of wheat (Triticum aestivum L.). However, the mutation rate via genome editing must be improved in order to analyze gene function and to quickly modify agronomic traits in wheat. Here, we examined the Cas9-induced mutation rates in wheat plants using two promoters for single guide RNA (sgRNA) expression and applying heat treatment during Agrobacterium tumefaciens-mediated transformation. Using the TaU6 promoter instead of the OsU6 promoter from rice (Oryza sativa L.) to drive sgRNA expression greatly improved the Cas9-induced mutation rate. Moreover, a heat treatment of 30°C for 1 day during tissue culture increased the Cas9-induced mutation rate and the variety of mutations obtained compared to tissue culture at the normal temperature (25°C). The same heat treatment did not affect the regeneration rates of transgenic plants but tended to increase the number of transgene integration sites in each transgenic plant. These results lay the foundation for improving the Cas9-induced mutation rate in wheat to enhance research on gene function and crop improvement.</p>
Journal
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- Plant Biotechnology
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Plant Biotechnology 40 (3), 237-245, 2023-09-25
Japanese Society for Plant Biotechnology
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Details 詳細情報について
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- CRID
- 1390297582102702080
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- NII Book ID
- AA11250821
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- ISSN
- 13476114
- 13424580
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- NDL BIB ID
- 033156624
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
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- Abstract License Flag
- Disallowed