Abstract
The CRISPR/Cas9 technique for rice genome engineering is gaining momentum and requires a precise gene delivery system. For rice and other crop plants, Agrobacterium tumefaciens-mediated transformation (AMT) is considered a suitable gene transformation method. The AMT for indica-type rice is a challenge because it is less efficient in tissue culture response than japonica-type rice. Here is a protocol of the AMT method that we developed for IR64 variety which has been successfully tested in other popular indica-type rice varieties. We used embryogenic calli as explant and an empty gRNA-containing CRISPR/Cas9 vector with hptII (hygromycin phosphotransferase) gene for the transformation. This technique would speed up rice genome editing via CRISPR/Cas9 technology and facilitate to achieve varied application in the future.
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Acknowledgments
This work was supported by Department of Biotechnology (DBT), Government of India, under the research project of “Translational Research on Transgenic Rice (Sanction #BT/PR12656/COE/34/22/2015)” and under the award (Prof. Swapan K. Datta) of “Distinguished Biotechnology Research Professor (DBT)” at the University of Calcutta, Kolkata, India.
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Majumder, S., Datta, K., Datta, S.K. (2021). Agrobacterium tumefaciens-Mediated Transformation of Rice by Hygromycin Phosphotransferase (hptII) Gene Containing CRISPR/Cas9 Vector. In: Bandyopadhyay, A., Thilmony, R. (eds) Rice Genome Engineering and Gene Editing. Methods in Molecular Biology, vol 2238. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1068-8_5
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DOI: https://doi.org/10.1007/978-1-0716-1068-8_5
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