Using Directed Evolution to Srudy the Origins of Multicellular Development

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Grant R35GM138030 utilizes directed evolution to study the evolution of multicellularity in the 'snowflake yeast' model system. The proposed work involved using single cell RNA-seq to examine the origin of multiple cell types over ~5,000 generations of evolution. Snowflake yseat evolve three distinct cell states, with elevated gene expression related to ribosomal processes, cell wall biogenesis, and stress response and programmed cell death, respectively. During a recent internship, undergraduate Luis Felipe Cedeno Perez developed a method to measure real-time single-cell phenotypic data in developing snowflake yeast clusters via soft-agar immobilization and image segmentation. He used machine learning to construct cell fate maps. His internship will end in April, 2023. If funded, Luis Felipe's summer project will allow him to complete his project, by integrating differential gene/protein expression into his segmentation pipeline, allowing for the construction of cell fate lineage maps and examination of location-dependent gene expression during ontogeny. Luis Felipe will gain expertise in experimental procedures and computational approaches, working closely with a postdoc in the lab for in-lab mentoring as well as meeting regularly with the PI. He will also participate in activities to improve his project management, scientific communication, and teamwork skills. The supplement will enable Luis Felipe to finish his project and generate the data required for a first-author paper, making him competitive for top-tier PhD programs in biomedical research. If he joins the Ratcliff lab as a PhD student, the project will give him a head start on his PhD thesis research and make him more competitive for proposals that would fund his PhD research.