TY - JOUR
T1 - Broad-spectrum resistance against multiple PVY-strains by CRSIPR/Cas13 system in Solanum tuberosum crop
AU - Noureen, Azka
AU - Zuhaib Khan, Muhammad
AU - Amin, Imran
AU - Zainab, Tayyaba
AU - Ahmad, Nasim
AU - Haider, Sibtain
AU - Mansoor, Shahid
N1 - KAUST Repository Item: Exported on 2022-06-08
Acknowledgements: This research was funded by a research grant from Higher Education Commission (HEC-project# 7308) and National Centre for Industrial Biotechnology (NCIB) for establishment of emerging CRISPR/Cas’s techniques in tetraploid Solanum tuberosum. We are thankful to our Collaborator Prof. Magdy M. Mahfouz (King Abdullah University of Science and Technology) for providing Cas13a gene for conducting this study.
This publication acknowledges KAUST support, but has no KAUST affiliated authors.
PY - 2022/6/2
Y1 - 2022/6/2
N2 - Potato virus Y (PVY) is a deadly environmental constraint that damages productivity of potato (Solanum tuberosum) around the globe. One of the major challenges is to develop resistance against PVY. Emerging clustered regularly short palindromic repeat (CRISPR)/Cas systems have the potential to develop resistance against PVY. In the current research, CRISPR-Cas13 has been exploited to target multiple strains of PVYN, PVYO, and PVYNTN. Multiple genes PI, HC-Pro, P3, Cl1, Cl2, and VPg 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.
AB - Potato virus Y (PVY) is a deadly environmental constraint that damages productivity of potato (Solanum tuberosum) around the globe. One of the major challenges is to develop resistance against PVY. Emerging clustered regularly short palindromic repeat (CRISPR)/Cas systems have the potential to develop resistance against PVY. In the current research, CRISPR-Cas13 has been exploited to target multiple strains of PVYN, PVYO, and PVYNTN. Multiple genes PI, HC-Pro, P3, Cl1, Cl2, and VPg 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.
UR - http://hdl.handle.net/10754/678713
UR - https://www.tandfonline.com/doi/full/10.1080/21645698.2022.2080481
U2 - 10.1080/21645698.2022.2080481
DO - 10.1080/21645698.2022.2080481
M3 - Article
C2 - 35652435
SN - 2164-5698
VL - 13
SP - 97
EP - 111
JO - GM crops & food
JF - GM crops & food
IS - 1
ER -