CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus Y
Potato (Solanum tuberosum L.) is an important staple food around the world, and potato virus Y (PVY) is a major constraint for potato production. The VPg protein of PVY interacts with the translation initiation factor eIF4E of the host that works as a susceptibility factor during infection. The inte...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-07-01
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| Series: | Frontiers in Genetics |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fgene.2022.922019/full |
| _version_ | 1797985366241705984 |
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| author | Azka Noureen Azka Noureen Muhammad Zuhaib Khan Imran Amin Tayyaba Zainab Tayyaba Zainab Shahid Mansoor |
| author_facet | Azka Noureen Azka Noureen Muhammad Zuhaib Khan Imran Amin Tayyaba Zainab Tayyaba Zainab Shahid Mansoor |
| author_sort | Azka Noureen |
| collection | DOAJ |
| description | Potato (Solanum tuberosum L.) is an important staple food around the world, and potato virus Y (PVY) is a major constraint for potato production. The VPg protein of PVY interacts with the translation initiation factor eIF4E of the host that works as a susceptibility factor during infection. The interaction between eIF4E and VPg was disrupted by CRISPR/Cas9. The homozygous conserved region of eIF4E of the potato variety “Kruda” was mutated by CRISPR/Cas9. Tracking of insertion, deletion, and conversion events was performed by Sanger sequencing with ∼15% editing efficiency. Truncated and mutated eIF4E proteins were unable to interact with VPg, and the virus was not able to exploit the host machinery for replication and systemic spreading. Mutated eIF4E lines showed enhanced resistance to PVYO strain. DAS-ELISA and RT-PCR were used for validation of the observed resistance. PVY resistance in tetraploid lines via CRISPR/Cas9 provides a route to develop novel resistant potato cultivars. |
| first_indexed | 2024-04-11T07:17:03Z |
| format | Article |
| id | doaj.art-ad1e1d760ebf4953b62cb5a977d0a83a |
| institution | Directory Open Access Journal |
| issn | 1664-8021 |
| language | English |
| last_indexed | 2024-04-11T07:17:03Z |
| publishDate | 2022-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Genetics |
| spelling | doaj.art-ad1e1d760ebf4953b62cb5a977d0a83a2022-12-22T04:37:56ZengFrontiers Media S.A.Frontiers in Genetics1664-80212022-07-011310.3389/fgene.2022.922019922019CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus YAzka Noureen0Azka Noureen1Muhammad Zuhaib Khan2Imran Amin3Tayyaba Zainab4Tayyaba Zainab5Shahid Mansoor6Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Pakistan Institute of Engineering and Applied Sciences (PIEAS), Faisalabad, PakistanUniversity Institute of Biochemistry and Biotechnology (UIBB), Pir Mehr Ali Shah-Arid Agriculture University, Rawalpindi, PakistanAgricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Pakistan Institute of Engineering and Applied Sciences (PIEAS), Faisalabad, PakistanAgricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Pakistan Institute of Engineering and Applied Sciences (PIEAS), Faisalabad, PakistanUniversity Institute of Biochemistry and Biotechnology (UIBB), Pir Mehr Ali Shah-Arid Agriculture University, Rawalpindi, PakistanNational Centre of Industrial Biotechnology (NCIB), Pir Mehr Ali Shah-Arid Agriculture University, Rawalpindi, PakistanAgricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Pakistan Institute of Engineering and Applied Sciences (PIEAS), Faisalabad, PakistanPotato (Solanum tuberosum L.) is an important staple food around the world, and potato virus Y (PVY) is a major constraint for potato production. The VPg protein of PVY interacts with the translation initiation factor eIF4E of the host that works as a susceptibility factor during infection. The interaction between eIF4E and VPg was disrupted by CRISPR/Cas9. The homozygous conserved region of eIF4E of the potato variety “Kruda” was mutated by CRISPR/Cas9. Tracking of insertion, deletion, and conversion events was performed by Sanger sequencing with ∼15% editing efficiency. Truncated and mutated eIF4E proteins were unable to interact with VPg, and the virus was not able to exploit the host machinery for replication and systemic spreading. Mutated eIF4E lines showed enhanced resistance to PVYO strain. DAS-ELISA and RT-PCR were used for validation of the observed resistance. PVY resistance in tetraploid lines via CRISPR/Cas9 provides a route to develop novel resistant potato cultivars.https://www.frontiersin.org/articles/10.3389/fgene.2022.922019/fullpotato virusesCRISPR-Cas9eIF4EPVYDAS-ELISART-PCR |
| spellingShingle | Azka Noureen Azka Noureen Muhammad Zuhaib Khan Imran Amin Tayyaba Zainab Tayyaba Zainab Shahid Mansoor CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus Y Frontiers in Genetics potato viruses CRISPR-Cas9 eIF4E PVY DAS-ELISA RT-PCR |
| title | CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus Y |
| title_full | CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus Y |
| title_fullStr | CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus Y |
| title_full_unstemmed | CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus Y |
| title_short | CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus Y |
| title_sort | crispr cas9 mediated targeting of susceptibility factor eif4e enhanced resistance against potato virus y |
| topic | potato viruses CRISPR-Cas9 eIF4E PVY DAS-ELISA RT-PCR |
| url | https://www.frontiersin.org/articles/10.3389/fgene.2022.922019/full |
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