Unlocking Nature’s Shield: The Promising Potential of CRISPRa in Amplifying Antimicrobial Peptide Expression in Common Bean (<i>Phaseolus vulgaris</i> L.)

This study proposes using the CRISPR transcriptional activation strategy to modulate the expression of genes encoding defense proteins and antimicrobial peptides (AMPs) in <i>Phaseolus vulgaris</i>. Three genes (PvD1, Pv-thionin, and Pv-lectin) were selected and targeted by the CRISPR-dCas9-TV-mediated transcriptional activation complex in the <i>P. vulgaris</i> L. hairy root. RT-qPCR investigated their activation efficiency. The eGFP-positive transgenic hairy roots exhibit enhanced expression of targeted genes compared to that of control roots. A moderate increase of 1.37-fold in PvD1 gene expression was observed in transgenic hairy roots, while 6.97-fold (Pv-lectin) and 5.70-fold (Pv-thionin) increases were observed. Importantly, no off-target effects of sgRNAs were detected, ensuring the precision and safety of the CRISPR-dCas9-TV strategy. The present article is a proof-of-concept study, and it has succeeded in demonstrating the efficiency of the CRISPR-dCas9-TV strategy in modulating the expression of target genes in <i>P. vulgaris</i>, paving the way for an alternative approach to protecting such essential crop plants.