Utilizing Target Sequences with Multiple Flanking Protospacer Adjacent Motif (PAM) Sites Reduces Off-Target Effects of the Cas9 Enzyme in Pineapple
We found that target sequences with multiple flanking PAM sites are more likely to bind with the Cas9 protein and induce mutations. Selecting sequences with multiple flanking PAM sites as editing targets can reduce the off-target effects of the Cas9 enzyme in pineapple. These findings provide a foundation for improving off-target prediction and engineering CRISPR-Cas9 complexes for gene editing.
Authors
- Chengjie Chen (ORCID: https://orcid.org/0000-0001-5964-604X)
- Qing Wei (ORCID: https://orcid.org/0000-0001-7803-1139)
- Hidayat Ullah (ORCID: https://orcid.org/0000-0002-2295-6623)
- Aiping Luan (ORCID: https://orcid.org/0000-0003-3774-2780)
- Rulin Zhan (ORCID: https://orcid.org/0000-0002-7561-4301)
- Haiyan Shu
- You Wang (ORCID: https://orcid.org/0000-0001-6058-9900)
- Junhu He
- Shenghe Chang
Institutions
- Chinese Academy of Tropical Agricultural Sciences (CN)
- Tropical Crops Genetic Resources Institute (CN)
- University of Swabi (PK)
Publication Details
- Journal
- Genes
- Published
- 2025-02-13
- DOI
- https://doi.org/10.3390/genes16020217
- Citations
- 1
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 0.81
Funders
- National Natural Science Foundation of China
- Chinese Academy of Tropical Agricultural Sciences
- Central Public-interest Scientific Institution Basal Research Fund for Chinese Academy of Tropical Agricultural Sciences