Broad-spectrum resistance against multiple PVY-strains by CRSIPR/Cas13 system in <i>Solanum tuberosum</i> crop
Azka Noureen, Muhammad Zuhaib Khan, Imran Amin, Tayyaba Zainab, Nasim Ahmad, Sibtain Haider, Shahid Mansoor
Abstract
genes of PVY were targeted by CRISPR/Cas13a. Multiplex gRNA cassettes were developed on the conserved regions of the PVY-genes. Three independent CRISPR/Cas13 transgenic potato lines were developed by applying an optimized concentration of trans-ribo zeatin and indole acetic acid at callus development, rooting, and shooting growth stages. The level of resistance in transgenic plants was confirmed through double-antibody sandwich enzyme-linked immunosorbent assay and real-time quantitative PCR. Our results have shown that efficiency of PVY inhibition was positively correlated with the Cas13a/sgRNA expression. Finding provides the specific functionality of Cas13 with specific gRNA cassette and engineering the potential resistance in potato crop against multiple strains of PVY.