Broad-spectrum resistance against multiple PVY-strains by CRSIPR/Cas13 system in Solanum tuberosum crop

Azka Noureen; Muhammad Zuhaib Khan; Imran Amin; Tayyaba Zainab; Nasim Ahmad; Sibtain Haider; Shahid Mansoor

doi:10.1080/21645698.2022.2080481

Broad-spectrum resistance against multiple PVY-strains by CRSIPR/Cas13 system in Solanum tuberosum crop

Azka Noureen, Muhammad Zuhaib Khan, Imran Amin, Tayyaba Zainab, Nasim Ahmad, Sibtain Haider, Shahid Mansoor

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	97-111
Number of pages	15
Journal	GM crops & food
Volume	13
Issue number	1
DOIs	https://doi.org/10.1080/21645698.2022.2080481
State	Published - Jun 2 2022
Externally published	Yes

Bibliographical note

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.

ASJC Scopus subject areas

General Medicine

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10.1080/21645698.2022.2080481

Cite this

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title = "Broad-spectrum resistance against multiple PVY-strains by CRSIPR/Cas13 system in Solanum tuberosum crop",

abstract = "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.",

author = "Azka Noureen and {Zuhaib Khan}, Muhammad and Imran Amin and Tayyaba Zainab and Nasim Ahmad and Sibtain Haider and Shahid Mansoor",

note = "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{\textquoteright}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.",

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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.

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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.

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ER -

Broad-spectrum resistance against multiple PVY-strains by CRSIPR/Cas13 system in Solanum tuberosum crop

Abstract

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