Develop and Optimize CRISPR Editing Tools for Biofuel Plant, Sorghum bicolor
Peggy G. Lemaux, Professor of Cooperative Extension
Plant and Microbial Biology
Closed. This professor is continuing with Spring 2024 apprentices on this project; no new apprentices needed for Fall 2024.
Sorghum (Sorghum bicolor L. Moench), is the fifth most important cereal crop worldwide and is a critical food, forage, and emerging biofuel crop. Understanding the photosynthetic mechanisms by which sorghum can capture sunlight more efficiently under adverse climate conditions is critical to using this crop to remove carbon dioxide that is increasing in levels in the atmosphere. For a Chan Zuckerberg Initiative-funded project, we are attempting to develop sorghum that will more effectively sequester additional carbon by identifying diverse gene targets involved in processes like optimizing photosynthesis, partitioning increased photosynthate to the roots, recovering faster from photodamage and making larger and deeper roots. Pioneering work from our team demonstrated more efficient transformation of sorghum by introducing transcriptional regulators, WUSCHEL2 (WUS2) and BABYBOOM (BBM), that reset the developmental pathways in transformed cells of the immature embryo. This expanded the number of transformable sorghum varieties and increased efficiency of introducing editing machinery.
Despite these advances, genome editing occurs at a low efficiency in sorghum, posing a barrier to using engineering approaches to study gene function. We are building on that capacity by applying new technical advances, with researchers at the Innovative Genomics Institute, to develop high-efficiency genome editing protocols. Current efforts focus on developing improved genome editing tools, including CRISPR/Cas9 editing, base editing, and prime editing. Genome editors will be optimized using sorghum protoplasts and then translating that into stable transformation efforts. Development of proper constructs with editor genes and guide RNAs and introduction into sorghum began in 2022. Once putative sorghum seedlings are generated, PCR will be used to determine the presence of gRNAs and the success in modifying target genes. Once confirmed, analyses will be done to assess effects of knock outs, knock ins or nucleotide modifications of the target genes on phenotypic, biochemical and molecular characteristics of the edited plants.
Role: Interests and skills of the student will dictate the specific duties in which the student will be involved. The student will first learn basic techniques used in the lab, which could include molecular cloning, gRNA design, sorghum protoplast transfection, sorghum transformation and molecular analyses of transgenic plants. Student will work with postdocs, research assistants, other undergraduates and the principal investigator and will participate in lab meetings and contribute to presentations. Time commitment will be negotiated.
Qualifications: Enthusiasm for research is required; some laboratory experience outside of the classroom is desired. Ability to work easily with a diverse laboratory group is also desired. Junior student is preferred.
Day-to-day supervisor for this project: Jianqiang Shen, Post-Doc
Hours: to be negotiated
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Biological & Health Sciences