A modular cloning toolkit for genome editing in plants (bioRxiv)

Genome editing with the CRISPR/Cas system is now widely used in functional studies across biological sciences including plant biology. Typically, this system involves a DNA nuclease and a guide RNA that directs the nuclease to a specific location in the genome. Golden Gate (GG) is a cloning method that is able to efficiently assemble multiple expression units and has been used successfully in plant genome editing applications. However, GG modules for some of the latest genome editing enzymes are lacking. Here, Hahn et al. have developed an additional cloning toolkit which contains ninety-nine modules encoding several CRISPR/Cas nucleases and their cognate guide RNA. In addition, the authors developed other genetic components including several promoters for driving the expression of CRISPR/Cas nuclease in monocots and dicots. Lastly, they also developed a set of modules that allows the construction of tRNA-sgRNA polycistronic units and demonstrated the application of this system in wheat. The resources are available from Addgene. (Summary by Toluwase Olukayode) bioRxiv 10.1101/738021