Here we also report a set of Gateway-based Cas9/sgRNA vectors that are highly efficient and easy to use for gene editing. The intermediate vector can be used to construct up to four sgRNA genes for single or multiplex gene editing in the rice genome. We demonstrated that most mutated To plants contain di-allelic (heterogeneous or homogeneous)modifications and that modified genes can be genetically segregated away from Cas9 and sgRNAtransgenes and faithfully transmitted through ensuing generations in a Mendelian fashion.We further demonstrated that deletions
of large chromosomal segments can be efficiently achieved in regenerated plants with our constructs. Together, these results suggested that the current set of Cas9/sgRNA constructs represents an important technical step forward that should be of significant benefit to the rice research community and agricultural industry in pursuing goals associated with customized genome editing and chromosome engineering.