Products/Services Used | Details | Operation |
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Gene Synthesis> | Overlapping gene fragments were synthesized from GenScript and cloned into expression cassettes containing the Zea mays Ubiquitin (UBI) promoter, 5′ untranslated region (UTR), and Intron 1 using the NEBuilder HiFi DNA Assembly kit (New England Biolabs) over BamHI and HpaI sites | Get A Quote |
CRISPR-Cas systems are robust and facile tools for manipulating the genome, epigenome and transcriptome of eukaryotic organisms. Most groups use class 2 effectors, such as Cas9 and Cas12a, however, other CRISPR-Cas systems may provide unique opportunities for genome engineering. Indeed, the multi-subunit composition of class 1 systems offers to expand the number of domains and functionalities that may be recruited to a genomic target. Here we report DNA targeting in Zea mays using a class 1 type I-E CRISPR-Cas system from S. thermophilus. First, we engineer its Cascade complex to modulate gene expression by tethering a plant transcriptional activation domain to 3 different subunits. Next, using an immunofluores... More