INVERT: crispr-based induction of chromosomal inversions in vegetable crops

Overview

INVERT is a genome-editing platform that enables the targeted induction of chromosomal inversions in vegetable crops using CRISPR/Cas technology. Chromosomal inversions are powerful tools for crop improvement because they can reshape genetic linkage, suppress undesirable recombination, and create novel trait combinations. INVERT addresses the central challenge that inversion frequencies in plants are typically very low and decrease as inversion size increases. The platform combines optimized guide RNA design, customized Cas variants, strategic microhomology placement, and species-tailored transformation methods to deliver transgene-free, stably inherited inversions.

Technical specifications

Key features and methods:

• Precision guide RNA and Cas engineering for accurate cleavage and reduced off-target effects, using ribonucleoprotein complexes or customized DNA constructs
• DNA repair pathway redirection toward microhomology-mediated end joining (MMEJ) through strategic placement of microhomologies at target sites
• High-efficiency transformation protocols adapted to each crop, including protoplast transfection and particle bombardment of reproductive tissues
• Marker-free, cisgenic plant generation with established transformation rates of approximately five percent
• Multiplexed editing capability using diverse CRISPR/Cas systems demonstrated across multiple plant families including Solanaceae
• Comprehensive method portfolio spanning non-targeted approaches such as TILLING and EcoTILLING alongside targeted editing tools including CRISPR and TALEN

Technology readiness level

The underlying CRISPR/Cas components have been validated in model and crop species including Arabidopsis and maize, where small to large inversions have been induced but at low frequency. The team has demonstrated functional genome-editing workflows, protoplast-based transformation, and mutation induction across several plant families. Remaining development focuses on systematically increasing inversion frequency and inheritance rates through combined optimization of Cas activity, MMEJ-directed repair, and species-specific delivery methods. The platform is positioned for collaborative pilot studies and applied validation in target vegetable crops.