Transgene-free genome editing in plants using rna-based viral delivery systems
A novel approach to plant genome editing that uses miniature CRISPR systems (Cas12j and Cas12f) delivered via (+)RNA viruses to produce non-GM, edited plants without genetic transformation. This method overcomes the size limitations of conventional CRISPR-Cas9 delivery, enabling systemic editing and heritable changes through viral mobility.
Overview
This research introduces a method for transgene-free genome editing in plants by leveraging miniature CRISPR systems delivered through RNA-based viral vectors. Conventional CRISPR-Cas9 systems are too large for effective viral delivery, limiting their use in transformation-free editing. By using compact Cas12j and Cas12f variants, this approach enables systemic genome editing throughout an entire plant without integrating foreign DNA into the plant genome. The result is non-genetically modified (non-GM) edited plants and potentially edited seeds, offering a significant advancement for plant biotechnology and breeding programs.
Technical specifications
Key features:
- Miniature CRISPR systems: Cas12j-2, CasMINI, AsCas12f1, and SpCas12f1, all under 2kb in size, enabling packaging within size-restricted viral genomes
- Viral delivery platform: Potato virus X (PVX) and other (+)RNA viruses engineered to carry CRISPR cassettes and guide RNA sequences
- Geminiviral replicon (GVR) system: Provides 10-50 times higher protein expression compared to binary plasmids for initial validation
- Plant codon optimization: CRISPR components synthesized with plant-preferred codons for enhanced expression in plant cells
- Meristematic targeting: Potential to produce heritable, transgene-free edits by infecting meristematic tissues, yielding edited seeds
- Scalable pathway: Designed for translation from model species (Nicotiana benthamiana) to monocot crops using viruses such as BSMV and FoMV
Technology readiness level
The technology is currently at an early-to-mid stage of development. The miniature CRISPR systems have been validated for genome editing activity in mammalian and plant systems, and the research team has designed and synthesized plant codon-optimized versions of five CRISPR cassettes. Initial validation will use bean yellow dwarf geminiviral replicons in transient assays in Nicotiana benthamiana, followed by PVX-based systemic infection studies. Future work includes evaluating heritable editing efficiency and extending the approach to monocot crops. The approach remains in the research and proof-of-concept phase, with further validation needed before commercial application.
About The University of Queensland
The University of Queensland is a large, multi-campus public research university in Brisbane with a comprehensive academic and research profile. Industry partners engage through hospital-embedded laboratories and clinical sites, pilot testbeds, and field campuses across Queensland that enable prototyping, trials, and validation in real-world settings. Co-location with major health precincts and proximity to Brisbane’s innovation districts make collaboration and talent access straightforward, while on-campus incubation and professional education support workforce upskilling. Research is supported by competitive national funding from the Australian Research Council and the National Health and Medical Research Council, alongside state and industry investment. A dedicated technology transfer office and commercialization company manage IP, licensing, sponsored research agreements, and spin-out formation.
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