TRANSFORM-PGR: Mining the compact Utricularia genome as source of novel regulatory elements for crop biotechnology

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The goal of this project is to advance plant genetic engineering technology and enable next-generation research and plant-improvement to help meet the upcoming challenges in agriculture. The need for increased but sustainable food production needs to be addressed through many approaches. Bioengineering can help address human food security through crop improvement. These improved crops will range from varieties highly adapted to diverse climates to crops with improved nutrition, that are better able to resist pests, use less water and fertilizer, or make the raw materials for bioplastics and other biodegradable materials. To make these plants a reality, it is first necessary to develop the ability to bioengineer multiple traits into plants. Current technology can place many genes close together and insert them into a crop; however, placing these genes close to each other causes them to interfere with each other. Hence, plants typically put a lot of space between their genes, but there is at least one plant known, the bladderwort, that has solved the problem of gene spacing its genome has many genes that are close together, but do not interfere with one another. The bladderwort also contains shortened versions of other DNA sequences that control when and where genes are expressed. This project uses bioinformatics to identify the bladderwort DNA sequences that allow for close spacing of genes, and to isolate the shorter versions of DNA control sequences. The function of each sequence will be validated empirically, and then made publicly available. The research team will also use the bladderwort sequences to find similar control sequences in other plants, including important crop species.Currently, the lack of appropriate DNA regulatory elements can make it difficult to get coordinated expression of multiple genes cloned into one vector. A few DNA insulators have been identified in plants that prevent interference from neighboring genes, but these insulators are so large their use makes vectors impossibly large. The compact genome (just 82 Mb) of Utricularia gibba (bladderwort) has solved many of these expression issues as it evolved. Expression patterns makes it clear this species has compact promoters, terminators, and insulators, as well as bidirectional promoters. Identification, cloning and validation of these elements and their function will provide a set of tools that will enable the routine use of multiple gene engineering. The DNA elements identified in this project will be made publicly available via AddGene, and thus contribute to a new generation of genetic engineering vectors. The researchers will also use the bladderwort sequences to identify similar conserved regulatory elements across angiosperms.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.