This project aims to develop enabling technology for studying essential gene functions in plant mitochondria. In plant cells, like in animals, mitochondria are known as the cellular powerhouses, generating the energy needed for essential activities. Mitochondria contain their own genomes, which are known to be more complex and much larger in plants than in animals; yet, how plant genomes work to produce characteristic RNA and protein products is not well understood. Moreover, genetic tools for studying genes in mitochondria are lacking. Therefore, the tools to be generated by this project could revolutionize the ability to analyze and potentially engineer mitochondrial genes and their products. In addition to this scientific impact, the project will have beneficial educational outcomes by providing undergraduate students with hands-on research training to prepare them to become members of the future STEM workforce.
This project aims to resolve the lack of genetic tools to probe mitochondrial gene functions by providing a technology that will allow editing of mitochondrial transcripts at certain nucleotides. The CRISPR revolution in genome editing relies on the use of nucleases, Cas proteins, which act in bacterial immunity by generating double-stranded breaks in invading phage DNAs. Recently, "dead" Cas proteins that have lost their endonuclease activity but retained their binding affinity for nucleic acids have been used to engineer nuclear single base editors acting on either DNA or RNA. This project aims to develop a similar tool for mitochondrial RNA. The impact of this work will provide a novel technology for the field of plant mitochondrial biology. It will open up unprecedented opportunities to understand the function of mitochondrial genes, gene regulatory sequences, and proteins by making precise changes that cannot be made by mutating nuclear genes that encode mitochondrial proteins. This methodology could also provide a way to genetically engineer plant mitochondrial function.
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.
