Crispr-based gene editing service for creating targeted chromosomal inversions in vegetable crops

Overview

Geneshifters, LLC provides a CRISPR-based gene editing service designed to generate targeted chromosomal inversions in vegetable crops without producing genetically modified organisms. The approach uses a dual sgRNA strategy delivered through an in-house optimized Agrobacterium-mediated plant transformation system, offering an alternative to particle bombardment that minimizes off-target DNA modifications. The result is a pathway to commercially viable, non-GMO gene-edited plants that can be advanced through conventional breeding pipelines.

Technical specifications

• Dual sgRNA CRISPR-Cas9 design to induce targeted gene or chromosomal inversions, significantly reducing the risk of off-target modifications
• Optimized Agrobacterium-mediated plant transformation with 50–80% efficiency across multiple crops including wheat, rice, canola, barley, chickpea, and maize, and adaptable to vegetable crops of the partner's choosing
• Crop- and variety-independent transformation system that produces stable transformants following simple Mendelian segregation, enabling easier recovery of non-GMO plants
• Bioinformatics-guided sgRNA selection to rule out potential off-target changes before editing
• Junction PCR amplification with primers designed around predicted double-strand breaks, followed by Sanger sequencing to confirm inversion events
• Inverse-PCR approach to identify plants carrying introgressions in unique genomic regions
• PCR-based marker system for marker-assisted breeding of the target inversion into diverse genetic backgrounds
• Quantitative PCR analysis to evaluate transcript abundance and splicing patterns of genes within the inverted segment compared to control plants
• T1 progeny screening to confirm stability of the inversion and select non-GMO segregants

Technology readiness level

The service is built on a validated Agrobacterium-mediated transformation platform that has been demonstrated across multiple crop species with 50–80% transformation efficiency. The dual sgRNA design and PCR-based validation workflows have been refined through prior gene-editing projects in other crops. Future validation will focus on confirming inversion events through junction PCR and Sanger sequencing, assessing gene expression changes via quantitative PCR, and verifying inheritance and non-GMO status in the T1 generation. The offering is positioned as a first-generation service ready for partner-sponsored projects and collaborative development.