Efficient Assembly of Large Multiplex CRISPR-Cas9 Guide Arrays for Maize Genome Editing
June 20, 2019| |
Genome editing in maize using CRISPR-Cas9 is an effective technique and researchers aim to target several genes at the same time. Genetic transformation of maize is expensive and requires a lot of work, thus, it would be beneficial for researchers to have vectors that target multiple genomic loci from one transformation event. One of the techniques to achieve this is to arrange Cas9 guides into a multiplex array that is transformed as a single locus to the plant. However, these arrays are difficult to create using traditional assembly methods. Researchers from the University of Massachusetts Amherst, USA, developed the MoClo system, which is an elaboration of Golden Gate cloning and tailor-fit for assembling larger multiplexed Cas9 guide arrays.
The system is detailed in Bio-101, an online database of life science protocols.
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