ABSTRACT
This chapter presents the technical advances aiming at the generation of insertion-based genome edits in plants using CRISPR/Cas9 systems. Together with the development of genome sequencing technologies, such as next generation sequencing (NGS) and third generation sequencing (TGS), it has been possible to fully sequence and decipher various plant genomes. In rice, large chromosomal excisions from 115 kb to 245 kb have been performed in protoplasts using CRISPR/Cas9 and the deletion of two clusters has been verified in regenerated T0 generation plants. Recently, in an extensive study, efficient inversions were formed in Arabidopsis, of which the inversions of up to 18 kb were successfully transmitted to the next generation driven by an egg cell-specific promoter for Cas9 expression. For plasmids with only the left homology arm, in 71 transformed plants, PCR-restriction enzyme (RE) and sequencing analyses were performed. The essence of plant breeding is to introduce or pyramid favorable traits (genes) and delete or knock-out unfavorable ones through various techniques.
