Synthetic biology platform for high-throughput genetic engineering of sorghum
A sorghum transformation platform enabling fast, high-throughput testing of gene constructs, Cas9/gRNA sequences, and promoter/UTR edits to accelerate breeding for improved photosynthesis, biomass, and grain yield in sorghum and other C4 crops.
Overview
This platform addresses a critical bottleneck in sorghum improvement: the high cost and long lead time of obtaining stable transformants. By enabling fast, high-throughput assessment of gene constructs in tissue culture, the platform accelerates the breeding cycle and expands the sorghum transformation toolbox. The research targets the petC gene, which encodes the Rieske FeS protein—a key component of the photosynthetic electron transport chain. Increasing Rieske content has been shown to enhance photosynthesis, biomass, and grain yield. The platform supports both transgenic approaches and CRISPR/Cas9-mediated gene editing, including targeted modifications to promoter and untranslated regions (UTRs) that can boost gene transcription and translation without introducing transgenic material.
Technical specifications
Core capabilities:
- High-throughput sorghum tissue culture transformation using an established Agrobacterium method
- Testing of multiple Cas9 variants and gRNA sequences for targeted and random edits in petC promoter and UTRs
- Golden Gate assembly of gene constructs, including a synthetic carotenoid pathway for visual assessment of transformation success
- Genomic DNA sequencing of tissue culture to confirm gene edits
- Measurement of petC transcript levels and Rieske protein abundance in de-etiolated tissue culture
- Incorporation of Baby Boom and Wuschel genes to facilitate introduction of genetic changes into commercial sorghum lines
Scientific foundation:
- Demonstrated that overexpressing petC enhances photosynthesis in the model C4 plant Setaria viridis
- Confirmed that transgenic sorghum with elevated Rieske protein shows higher biomass and grain yield under glasshouse conditions
- Established synthetic metabolic pathway testing in transformed rice tissue culture
- Successfully applied CRISPR/Cas9 gene editing in Setaria viridis
Technology readiness level
The platform builds on validated proof-of-concept work in both Setaria viridis and sorghum. Transgenic sorghum overexpressing petC has been confirmed to deliver increased biomass and grain yield in glasshouse trials. CRISPR/Cas9 editing has been successfully demonstrated in Setaria viridis, and synthetic metabolic pathway testing has been established in rice tissue culture. The next phase involves transforming Sorghum bicolor Tx430 tissue culture, testing diverse Cas9/gRNA combinations, and evaluating edits in petC regulatory regions. This work aims to identify new gene elements that confer enhanced Rieske content through non-transgenic breeding approaches applicable to commercial sorghum varieties and other C4 crops.
About Monash University, Melbourne
Monash University is a comprehensive public research university and one of Australia’s largest, known for scale, interdisciplinarity, and an applied orientation. Its Melbourne-based technology precinct brings together university laboratories, pilot-scale and prototyping suites, and company R&D groups alongside government research organizations to enable co-development and rapid iteration. Integration with a major hospital network supports clinical trials and translation, while structured industry placements and doctoral partnerships create a robust talent pipeline for corporate R&D. Research is backed by competitive funding from the Australian Research Council, the National Health and Medical Research Council, and state and federal programs that incentivize industry collaboration. A dedicated technology transfer office manages IP, licensing, and startup formation, with pathways to incubation and investment within the precinct.
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