Transgene-free gene-edited vegetables via biolistic delivery platform

Overview

GRA&GREEN Inc. has developed an in-house biolistic gene-editing platform capable of producing transgene-free edited vegetables across multiple crop families, including tomato, watermelon, and pumpkin. The approach uses particle bombardment to deliver gene-editing tools into plant tissue without integrating foreign DNA into the genome, resulting in edited crops that may face fewer regulatory hurdles compared to transgenic alternatives. The company is seeking collaboration to advance the platform toward large-scale gene-editing screens, particularly targeting 2 Mbp chromosomal inversions, by combining biolistic delivery with morphogenetic factors that enhance regeneration efficiency in easily scalable explants.

Technical specifications

Key features:

• Biolistic transient expression system validated for transgene-free gene editing in tomato, watermelon, and pumpkin
• Expertise in Solanaceae and Cucurbitaceae crop pipelines, including dedicated teams for biolistic delivery, mutation detection, and seed multiplication
• Preliminary validation showing morphogenetic factors can induce de novo multiple shoot formation in tomato, enabling recovery of multiple independent edited lines from a single biolistic event
• Planned parallel screening of gRNA efficiency using a protoplast system, including evaluation of Cas9 and Cas12a (Cpf1) for generating complementary overhangs that may increase inversion rates
• Scalable explant strategy targeting tissues such as leaf, stem, or stem node for high-throughput delivery
• Operates within approved plant handling facilities and holds a commercial CRISPR license

How it works:

Gene-editing reagents are introduced into plant cells using particle bombardment rather than Agrobacterium-mediated transformation. Because the editing machinery is delivered transiently and no plasmid or selectable marker is integrated, resulting plants contain only the intended genomic edits. Morphogenetic factors are co-delivered to promote regeneration of edited cells into whole plants, allowing large populations of independent edited lines to be recovered and screened for desired genotypes, including large-scale chromosomal inversions generated by paired gRNAs.

Technology readiness level

The core biolistic transgene-free editing pipeline has been validated in commercial crops (tomato, watermelon, pumpkin), and preliminary experiments have demonstrated that morphogenetic factors can drive multiple shoot regeneration from bombarded tomato tissue. The proposed collaboration focuses on advancing the platform from proof-of-concept toward a robust large-scale screening workflow, including gRNA pair validation in protoplasts and explant optimization. Further validation phases are planned to identify optimal tissue targets and editing conditions before scaling to full genome-level screening for commercially relevant traits.