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- Title
In silico Prediction and Analysis of Potential Off-Targets and Off-Target Mutation Detection in StERF3-Gene Edited Potato Plants.
- Authors
Razzaq, Hafiza Arooj; Ijaz, Siddra; Haq, Imran Ul; Awan, Faisal Saeed
- 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.
- Subjects
GENOME editing; GENETIC mutation; CRISPRS; ENDONUCLEASES; PLANT development
- Publication
Phyton (0031-9457), 2023, Vol 92, Issue 8, p2451
- ISSN
0031-9457
- Publication type
Article
- DOI
10.32604/phyton.2023.030501