Dissertation

Improving targeted gene editing in Arabidopsis thaliana

Understanding DNA repair mechanisms is essential for advancing plant biotechnology and crop improvement. This study investigates how double-strand breaks (DSBs), induced by CRISPR/Cas systems, are repaired in the model plant Arabidopsis thaliana, and how these processes influence Agrobacterium tumefaciens T-DNA integration.

Author

L. Kamoen

Date

10 June 2025

Links

Thesis in Leiden Repository

DSBs are primarily repaired via classical nonhomologous end joining (cNHEJ) and polymerase theta-mediated end joining (TMEJ), each leading to distinct mutational outcomes. T-DNA integration at induced DSB sites was found to be frequent, with TMEJ playing a key role in mediating attachment of the T-DNA ends. By combining CRISPR-induced DSBs with Agrobacterium-mediated transformation, this research demonstrates a powerful strategy for targeted gene insertion. Finally, a novel method for error-free gene editing is developed, bypassing the low efficiency of homologous recombination. This work enhances our understanding of CRISPR-mediated genome editing and supports the development of precise, efficient strategies for plant genetic engineering.

• crispr/cas
• dna repair
• double-stranded break repair
• gene editing
• microhomology
• non-homologous end-joining
• polymerase theta-mediated end-joining
• t-dna