Dna-free gene editing platform for vegetatively propagated crops
An innovative DNA-free gene editing approach for crops with asexual reproduction that avoids protoplast isolation. Combines plant developmental regulators with transient suppression of Agrobacterium T-DNA integration genes to produce transgene-free edited plants, enabling precision breeding in species where conventional transformation and seed segregation eliminate desired traits.
Overview
This technology addresses a critical bottleneck in crop biotechnology: gene editing of vegetatively propagated species that cannot produce true seeds. Traditional genome editing in these crops results in GMO products because there is no sexual reproduction pathway to eliminate transgenes. The platform enables DNA-free, transgene-free gene editing without the need for protoplast isolation and culture, which remains extremely challenging in most crop species.
The solution is particularly valuable for highly heterozygous crops where desirable traits are lost during seed segregation in conventional breeding. By enabling precise, DNA-free edits, the technology preserves complex trait combinations while introducing new characteristics such as disease resistance.
Technical specifications
Core methodology:
- Co-expression of plant developmental regulators, including PLT5, to enhance in-planta and in vitro regeneration across multiple plant species
- Transient silencing of conserved Agrobacterium genes (POLQ and MRE11) to prevent T-DNA integration into the host genome
- Application targeting the DMR6 gene, which confers broad-spectrum resistance to bacterial leaf spot diseases
- Avoids protoplast isolation, making it applicable to species recalcitrant to standard tissue culture protocols
Demonstrated capabilities:
- Improved genetic transformation efficiency in several plant species through developmental regulator manipulation
- Successful DNA-free gene editing in model species
- Provisional patents filed for both the developmental regulator approach and the T-DNA suppression method
Technology readiness level
The technology has been validated in model species with provisional patents filed for both component methods. Current efforts focus on extending the platform to non-model, vegetatively propagated crops, with an initial target on a highly heterozygous supertunia variety. The 16-month validation plan aims to demonstrate improved DNA-free editing of DMR6 alongside enhanced in vitro regeneration. Success is expected to translate readily to other economically important vegetatively propagated crops.
About University of Florida
The University of Florida is a comprehensive public research university with a broad academic and research portfolio and a statewide presence. Industry collaborates through co-located labs and shared core facilities and through an integrated academic health system that accelerates clinical translation. A statewide extension network and multiple research and education sites connect companies with field-scale testing and rapid deployment, while incubators and an adjacent innovation district provide pathways from lab to market. Research is supported by competitive funding from major federal agencies such as NIH, NSF, USDA, and DOE. A dedicated technology transfer office supports IP, licensing, and startup formation.
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