University of Massachusetts Amherst scientists developed an efficient assembly of large multiplex CRIPR-Cas9 guide arrays for Genome editing of maize. Details of the protocol are published in Bio-Protocol.
CRISPR-Cas9 has been known as an effective tool for genome editing, which makes many researchers wishes to use the tool for targeting multiple genes. To address this, researchers developed vectors that target multiple genomic loci from a single transformation event. One of the proposed systems is called MoClo, which is an elaboration of Golden Gate cloning and is particularly fit for assembling larger multiplexed Cas9 guide arrays.
Jarret Man and Madelaine Bartlett described the steps for designing and building a custom guide array targeting any number of maize loci using the MoClo standard components and syntax. Instructions for using variations of the maize and rice U6 to drive guide RNA expression were also provided.