High-efficiency HDR genome editing in vegetables using TiGER workflow

Overview

This offering provides crop developers and breeding programs with access to Hudson River Biotechnology's TiGER workflow, a precision genome editing platform purpose-built for vegetables. The service addresses one of the most persistent bottlenecks in plant biotechnology: achieving reliable homology-directed repair (HDR) at efficiencies high enough to produce edited plants without extensive screening. By combining optimized protoplast transfection with an automated single-cell regeneration pipeline, the platform routinely delivers HDR efficiencies above 15% across more than 15 plant species, including recalcitrant varieties within Solanaceae and Cucurbitaceae families.

The value proposition is straightforward: faster, more predictable generation of plants carrying precise, desired alleles rather than the random knockouts typical of standard CRISPR/Cas approaches. This enables trait development work that requires exact sequence changes, such as introducing or modifying specific alleles for disease resistance, quality traits, or metabolic pathway engineering.

Technical specifications

Core workflow components:

• TiGER editing pipeline: Protoplast-based delivery of ribonucleoprotein (RNP) complexes together with donor DNA templates, applied at carefully titrated concentrations to favor HDR over non-homologous end joining (NHEJ)
• HDR efficiency: Consistently greater than 15% across validated vegetable species, driven by high RNP and donor template concentrations within protoplasts
• Controllability: Editing parameters including nuclease choice (MAD7), donor template design, and temperature are tuned to maximize HDR rates
• Regeneration pipeline: Automated screening of media components, hormone combinations, and regeneration enhancers supports single-cell regeneration from more than 15 plant species, including historically difficult varieties

Project structure (1.5-year engagement):

• Phase 0: Establishing in vitro culture (1 to 3 months)
• Phase 1: HDR optimization and off-target assessment (up to 12 months)
• Phase 2: Mutant creation, including protoplast transfection, shoot and root induction, and evaluation by in-house Nanopore sequencing (6 months)

Deliverables include 3 to 5 independently edited lines, with go/no-go decision points integrated throughout. Activities run in parallel where feasible, and project start is flexible throughout the year.

Technology readiness level

The TiGER workflow has been validated across more than 15 plant species and is in active commercial use for vegetable genome editing projects. HDR efficiencies above 15% have been reproducibly demonstrated in Solanaceae and Cucurbitaceae crops. Each new engagement begins with a Phase 0 to Phase 1 optimization step to confirm performance in the specific target variety, followed by mutant generation in Phase 2. Timelines may shift depending on the regeneration time of the chosen species, and go/no-go checkpoints are built in to manage risk. Edited lines are delivered back ready for downstream product development.