Microparticle-mediated CRISPR DNA delivery for genome editing in poplar
- Author
- Lennart Hoengenaert (UGent) , Jan Van Doorsselaere (UGent) , Ruben Vanholme (UGent) and Wout Boerjan (UGent)
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- Project
- Abstract
- The use of CRISPR/Cas9 is currently the method of choice for precise genome engineering in plants, including in the biomass crop poplar. The most commonly used method for delivering CRISPR/Cas9 and its components in poplar is via Agrobacterium-mediated transformation, that besides the desired gene-editing event also results in stable T-DNA integration. Here we explore the delivery of the gene-editing reagents via DNA-coated microparticle bombardment into the model tree Populus tremula x P. alba to evaluate its potential for developing transgene-free, gene-edited trees, as well as its potential for integrating donor DNA at specific target sites. Using an optimized transformation method, which favors the regeneration of plants that transiently express the genes on the delivered donor DNA, we regenerated gene-edited plants that are free of the Cas9 and the antibiotic resistance-encoding transgenes. In addition, we report the frequent integration of donor DNA fragments at the Cas9-induced double-strand break, opening opportunities toward targeted gene insertions.
- Keywords
- CRISPR, gene editing, particle bombardment, poplar, targeted insertions, tissue culture, transgene-free, TARGETED MUTAGENESIS, EXPRESSION, PLANTS, TRANSFORMATION, FREQUENCIES, INTEGRATION, SALICACEAE, REDUCTASE
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Hoengenaert et al. (2023)Frontiers in Plant Science 14,1286663.pdf
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-01HF9K16PFYYPWPHQPVKKJFS6Q
- MLA
- Hoengenaert, Lennart, et al. “Microparticle-Mediated CRISPR DNA Delivery for Genome Editing in Poplar.” FRONTIERS IN PLANT SCIENCE, vol. 14, 2023, doi:10.3389/fpls.2023.1286663.
- APA
- Hoengenaert, L., Van Doorsselaere, J., Vanholme, R., & Boerjan, W. (2023). Microparticle-mediated CRISPR DNA delivery for genome editing in poplar. FRONTIERS IN PLANT SCIENCE, 14. https://doi.org/10.3389/fpls.2023.1286663
- Chicago author-date
- Hoengenaert, Lennart, Jan Van Doorsselaere, Ruben Vanholme, and Wout Boerjan. 2023. “Microparticle-Mediated CRISPR DNA Delivery for Genome Editing in Poplar.” FRONTIERS IN PLANT SCIENCE 14. https://doi.org/10.3389/fpls.2023.1286663.
- Chicago author-date (all authors)
- Hoengenaert, Lennart, Jan Van Doorsselaere, Ruben Vanholme, and Wout Boerjan. 2023. “Microparticle-Mediated CRISPR DNA Delivery for Genome Editing in Poplar.” FRONTIERS IN PLANT SCIENCE 14. doi:10.3389/fpls.2023.1286663.
- Vancouver
- 1.Hoengenaert L, Van Doorsselaere J, Vanholme R, Boerjan W. Microparticle-mediated CRISPR DNA delivery for genome editing in poplar. FRONTIERS IN PLANT SCIENCE. 2023;14.
- IEEE
- [1]L. Hoengenaert, J. Van Doorsselaere, R. Vanholme, and W. Boerjan, “Microparticle-mediated CRISPR DNA delivery for genome editing in poplar,” FRONTIERS IN PLANT SCIENCE, vol. 14, 2023.
@article{01HF9K16PFYYPWPHQPVKKJFS6Q, abstract = {{The use of CRISPR/Cas9 is currently the method of choice for precise genome engineering in plants, including in the biomass crop poplar. The most commonly used method for delivering CRISPR/Cas9 and its components in poplar is via Agrobacterium-mediated transformation, that besides the desired gene-editing event also results in stable T-DNA integration. Here we explore the delivery of the gene-editing reagents via DNA-coated microparticle bombardment into the model tree Populus tremula x P. alba to evaluate its potential for developing transgene-free, gene-edited trees, as well as its potential for integrating donor DNA at specific target sites. Using an optimized transformation method, which favors the regeneration of plants that transiently express the genes on the delivered donor DNA, we regenerated gene-edited plants that are free of the Cas9 and the antibiotic resistance-encoding transgenes. In addition, we report the frequent integration of donor DNA fragments at the Cas9-induced double-strand break, opening opportunities toward targeted gene insertions.}}, articleno = {{1286663}}, author = {{Hoengenaert, Lennart and Van Doorsselaere, Jan and Vanholme, Ruben and Boerjan, Wout}}, issn = {{1664-462X}}, journal = {{FRONTIERS IN PLANT SCIENCE}}, keywords = {{CRISPR,gene editing,particle bombardment,poplar,targeted insertions,tissue culture,transgene-free,TARGETED MUTAGENESIS,EXPRESSION,PLANTS,TRANSFORMATION,FREQUENCIES,INTEGRATION,SALICACEAE,REDUCTASE}}, language = {{eng}}, pages = {{13}}, title = {{Microparticle-mediated CRISPR DNA delivery for genome editing in poplar}}, url = {{http://doi.org/10.3389/fpls.2023.1286663}}, volume = {{14}}, year = {{2023}}, }
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