Database Research Summaries2018 Platform for Fast and Cost-efficient CRISPR gene
Research Focus
The focus of this project is to find cost-effective and sustainable solutions to fight pathogens, pests, and diverse environmental stresses.
Objectives
- Accelerate soybean gene editing by administering the CRISPR-guiding RNAs (gRNA) using a virus.
- Adopt Cpf1, a more versatile Cas9 analogue, for gene insertion in soybean.
Results
- Both the bean pod mottle virus (BPMV) and apple latent spherical virus (ALSV) were tested as delivery vehicles for CRISPR guide RNAs. Two gRNAs were inserted into BPMV, and tested for their ability to induce photobleaching in transgenic soybean expressing a Cas9 gene (thus making the Cas9protein). A small number of soybean leaf cells were found to undergo the expected gene editing. However, leaves did not become photobleached.
- Transgenic plants carrying the CPF1 transgene have been obtained for both the Jack and Williams 82 cultivars. Our lab in collaboration with John Finer lab at OARDC generated the Jack variety CPF1 plants. The Williams 82 variety CPF1 plants were generated by Wisconsin Crop innovation Center (WCIC). Currently genotyping experiments are being conducted to assess the expression of CPF1 in these plants, as well as the efficiency of CPF1-mediated gene editing.
Importance
- Successful use of a soybean virus (bean pod mottle virus, BPMV) as the vehicle for gRNA delivery would take advantage of existing transgenic soybean lines that express the Cas9 gene editing enzyme, and substantially shorten the time needed for generating soybean plants with desired traits via CRISPR editing.
- The optimized procedure could then be applied to quickly testing previously identified candidate genes for their yield-enhancing, stress-tolerant potentials.
- The adoption of the novel CRISPR editing enzyme Cpf1 would further simplify the design of guide RNA constructs, permitting simultaneous editing of multiple traits. This would allow for the pyramiding of multiple beneficial traits with one single manipulation, saving years of breeding time.
For more information about this research project, please visit the National Soybean Checkoff Research Database.
Funded in part by the soybean checkoff.
