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ARTICLE
In silico Prediction and Analysis of Potential Off-Targets and Off-Target Mutation Detection in StERF3-Gene Edited Potato Plants
Hafiza Arooj Razzaq1, Siddra Ijaz1,*, Imran Ul Haq2, Faisal Saeed Awan1
1
Centre of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture, Faisalabad, 38040, Pakistan
2
Department of Plant Pathology, University of Agriculture, Faisalabad, 38040, Pakistan
* Corresponding Author: Siddra Ijaz. Email:
(This article belongs to this Special Issue: Agricultural Intensification, Climate Change, and Food Security)
Phyton-International Journal of Experimental Botany 2023, 92(8), 2451-2460. https://doi.org/10.32604/phyton.2023.030501
Received 10 April 2023; Accepted 11 May 2023; Issue published 25 June 2023
Abstract
The imperative aspect of the CRISPR/Cas9 system is a short stretch of 20 nucleotides of gRNA that control the
overall specificity. Due to the small size, the chance of its multiple occurrences in the genome increases; however,
a few mismatches are tolerated by the Cas9 endonuclease activity. An accurate and careful in silico-based off-target prediction while target selection is preferred to address the issue. These predictions are based on a comprehensive set of selectable parameters. Therefore, we investigated the possible off-target prediction and their
screening in
StERF3 gene-edited potato plants while developing
StERF3-loss-of-function mutants using
CRISPR/Cas9 approach. The 201 off-targets for the selected targets of the
StERF3 gene were predicted, and
79 were filtered as potential off-targets. Of these 79, twenty-five off-targets showed scores with defined cut-off
values <0.5 and were analyzed in Sterf3-edited potato plants compared to wild-type plants. No off-targeting
was found to have occurred in edited plants.
Keywords
Supplementary Material
Supplementary Material File
Cite This Article
Razzaq, H. A., Ijaz, S., Haq, I. U., Awan, F. S. (2023).
In silico Prediction and Analysis of Potential Off-Targets and Off-Target Mutation Detection in
StERF3-Gene Edited Potato Plants.
Phyton-International Journal of Experimental Botany, 92(8), 2451–2460.