Versatile Applications of CRISPR-Based Programmable T-DNA Integration in Plants.

ABSTRACT: Agrobacterium-mediated T-DNA integration into plant genomes represents a cornerstone for transgenic expression in plant basic research and synthetic biology. However, random T-DNA integration can disrupt essential endogenous genes or compromise transgene expression, stressing the need for targeted integration strategies. Here we explored CRISPR-aided targeted T-DNA integration (CRISTTIN) in Arabidopsis, leveraging CRISPR-induced double-strand breaks (DSBs) to facilitate precise T-DNA insertion. Contrary to our initial hypothesis, conventional Cas9 outperformed a designed Cas9-adaptor fusion nuclease that may recruit Agrobacterium VirD2/T-DNA complexes to DSB sites via the adaptor-VirD2 interaction. Using Cas9-based CRISTTIN, we streamlined the parallel generation of FERONIA null alleles an

SUBMITTER: Lin XX 

PROVIDER: S-EPMC12665094 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature