dCas9-BE3 and dCas12a-BE3 Systems Mediated Base Editing in Kiwifruit Canker Causal Agent Pseudomonas syringae pv. actinidiae
Abstract
:1. Introduction
2. Result
2.1. Genome Information of PSA.AH.01
2.2. Successfully Knockout Flic in Psa
2.3. Editing Rule of dCas9-BE3 in Psa
2.4. Multi-Site Knockout by Using the dCas9-BE3 System
2.5. pdCas12a-BE Knockout System
2.6. Construction of sgRNA Libraries Targeting Genome-Wide Editing
2.7. Pathogenicity Assays of Psa Mutant Strains
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains and Plant Growth Conditions
4.2. Genome Sequencing and Annotation
4.3. Construction of the Base-Editing System pdCas12a-BE
4.4. Target Design Procedure
4.5. Knockout Vector Construction
4.6. Construction of a Genome-Wide Mutant Library
4.7. Plasmid Transformations into Psa
4.8. Pathogenicity Assays
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Liu, B.; Song, W.; Wang, L.; Wu, Y.; Xu, X.; Niu, X.; Huang, S.; Liu, Y.; Tang, W. dCas9-BE3 and dCas12a-BE3 Systems Mediated Base Editing in Kiwifruit Canker Causal Agent Pseudomonas syringae pv. actinidiae. Int. J. Mol. Sci. 2023, 24, 4597. https://doi.org/10.3390/ijms24054597
Liu B, Song W, Wang L, Wu Y, Xu X, Niu X, Huang S, Liu Y, Tang W. dCas9-BE3 and dCas12a-BE3 Systems Mediated Base Editing in Kiwifruit Canker Causal Agent Pseudomonas syringae pv. actinidiae. International Journal of Molecular Sciences. 2023; 24(5):4597. https://doi.org/10.3390/ijms24054597
Chicago/Turabian StyleLiu, Bo, Wenpeng Song, Linchao Wang, Yantao Wu, Xiaoting Xu, Xiangli Niu, Shengxiong Huang, Yongsheng Liu, and Wei Tang. 2023. "dCas9-BE3 and dCas12a-BE3 Systems Mediated Base Editing in Kiwifruit Canker Causal Agent Pseudomonas syringae pv. actinidiae" International Journal of Molecular Sciences 24, no. 5: 4597. https://doi.org/10.3390/ijms24054597