The great advances in exploiting the CRISPR/Cas9 system are paving the way for targeted genome engineering in plants. Genome editing by direct delivery of CRISPR/Cas9 ribonucleoprotein complexes (RNPs) into plant cells reduces off-target mutations and avoids the integration of foreign DNA fragments, thus providing an efficient and accurate method for precision crop breeding. Here we describe an RNP-based genome editing protocol for wheat. The protocol covers the in vitro transcription of sgRNA, purification of Cas9 protein, biolistic delivery of CRISPR/Cas9 RNPs, and tissue culture procedures for regenerating testable seedlings.
CITATION STYLE
Liang, Z., Chen, K., & Gao, C. (2019). Biolistic delivery of CRISPR/Cas9 with ribonucleoprotein complex in wheat. In Methods in Molecular Biology (Vol. 1917, pp. 327–335). Humana Press Inc. https://doi.org/10.1007/978-1-4939-8991-1_24
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