Efficient Assembly of Large Multiplex CRISPR-Cas9 Guide Arrays for Maize Genome Editing
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.
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bio-protocol.org]