Selenium Nanoparticle Platform for Tissue-Culture-Free CRISPR Donor Delivery in Plants

Overview

This offering is a selenium-nanoparticle (SeNP) based delivery platform for plant genome editing. It enables co-delivery of CRISPR-Cas machinery—either ribonucleoprotein (RNP) or plasmid—with donor DNA templates in a tissue-culture-free, topical application. The ~20 nm BSA-stabilized SeNP colloid acts as a non-viral nucleic acid carrier, avoiding the need for tissue culture and reducing the somaclonal variation often associated with conventional transformation routes. The platform is designed to address a key bottleneck in plant precision breeding: delivering large donor templates (up to 7–10 kb) alongside editing reagents to achieve precise, single-copy, heritable insertions via homology-directed repair (HDR).

Technical specifications

Core features:

• Non-viral, tissue-culture-free delivery via topical application to leaves or meristems
• ~20 nm BSA-stabilized selenium nanoparticles with colloidal stability
• Co-loading of CRISPR-Cas reagents (RNP or plasmid) and donor DNA in a single complex
• Compatible with both Cas9 and Cas12a editing systems
• Scalable loading from current reporter/Cas9-sized constructs toward 7–10 kb donor templates
• Characterization via DLS, SEM, TEM, XRD, and UV-Vis supports reproducibility

Validation and development path:

• Demonstrated nucleic-acid carrier performance in Arabidopsis, benchmarked against PEI-carbon nanotubes
• Active funded programs extend the platform to Cucurbita pepo (CRISPR/Cas9 editing via topical meristem/leaf application) and recalcitrant woody crops under Horizon Europe GENIUS
• Planned transfer to oilseed rape (Brassica napus), with soybean as a subsequent target pending material access
• Event quality (single-copy status, junction integrity, heritability) will be assessed through molecular analysis pipelines

Technology readiness level

The core SeNP carrier mechanism has been validated in Arabidopsis, placing the fundamental technology at approximately TRL 2–3 for large-payload editing applications. The current proposal targets advancement to TRL 3–5 over a 12–18 month sponsored research or co-development programme. Large-cassette knock-in data and validation in oilseed rape and soybean are not yet generated and are the explicit focus of this development effort. Collaboration is sought to contribute editing machinery, donor designs, germplasm, and advanced characterization to accelerate the path toward deployment.