The repurposing of type I-E CRISPR-Cascade for gene activation in plants

Communications Biology - Tập 2 Số 1
Joshua K. Young1, Stephen L. Gasior1, Spencer Jones1, Lijuan Wang1, Pedro Navarro1, Becca Vickroy1, Rodolphe Barrangou2
1Department of Molecular Engineering, Corteva Agriscience™, Johnston, IA, 50131, USA
2Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695, USA

Tóm tắt

AbstractCRISPR-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 immunofluorescent assay, we confirm Cascade cellular complex formation and observe enhanced gene activation when multiple subunits tagged with the transcriptional activator are combined. Finally, we examine Cascade mediated gene activation at chromosomal DNA targets by reprogramming Zea mays cells to change color.

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Communications Biology

Tập 2 Số 1

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