Generating mutants carrying multiple genetic alterations is essential for genetic pathway research and crop improvement. The conventional process generating multiple mutations is both time-consuming and labor intensive. Recently, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9) system is emerging as a powerful tool for genome editing in various organisms, including plants. The rapid development of CRISPR/Cas9 system provides a potential strategy for multiplex genome editing in plants.

Phenotypes of the transgenic rice plants

Recently, researchers from China National Rice Research Institute (CNRRI) and Yangzhou University developed a simple system to generate multiple mutations in rice by co-expression of Cas9 protein and multiple gRNAs targeting different genomic loci. By rapid construction of the vectors and a single transformation, multiple genes were mutated with high-efficiency. Further statistical analysis revealed that mutations tend to occur simultaneously and different targets showed different proportions of mutation types in the transgenic plants, indicating different sgRNAs function independently of each other. Thus, the establishment of CRISPR-Cas9 vector system allows convenient generation of rice carrying mutations in multiple genes, thereby facilitating both basic research and crop improvement.

Schematic diagram illustrating the structure of vectors and construction strategy

This work was supported by the Agricultural Science and Technology Innovation Program. The research finding has been published in Journal of Genetics and Genomics online on October 24th 2015 (DOI: doi:10.1016/j.jgg.2015.09.011). More details are available on the links bellow: