AUTHOR=Noureen Azka , Khan Muhammad Zuhaib , Amin Imran , Zainab Tayyaba , Mansoor Shahid 

TITLE=CRISPR/Cas9-Mediated Targeting of Susceptibility Factor eIF4E-Enhanced Resistance Against Potato Virus Y

JOURNAL=Frontiers in Genetics

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2022.922019

DOI=10.3389/fgene.2022.922019

ISSN=1664-8021

ABSTRACT=<p>Potato (<italic>Solanum tuberosum</italic> 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 <italic>eIF4E</italic> 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 <italic>eIF4E</italic> 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 <italic>eIF4E</italic> lines showed enhanced resistance to PVY<sup>O</sup> strain. DAS-ELISA and RT-PCR were used for validation of the observed resistance. PVY resistance in tetraploid lines <italic>via</italic> CRISPR/Cas9 provides a route to develop novel resistant potato cultivars.</p>