Using Directed Evolution to Srudy the Origins of Multicellular Development

格兰特R35GM138030利用定向进化研究雪花中多细胞的进化 酵母菌模型系统。拟议的工作涉及使用单细胞rna-seq来检查多个 细胞类型经过约5,000代的进化。雪花YSEAT进化出三种不同的细胞状态， 与核糖体过程、细胞壁生物发生、应激反应和程序化细胞相关的基因表达 分别是死亡。在最近的一次实习中，本科生路易斯·费利佩·塞德诺·佩雷斯开发了一种方法 软琼脂培养雪花酵母团时实时单细胞表型数据的测定 固定和图像分割。他使用机器学习来构建细胞命运图。他的实习经历 将于2023年4月结束。如果获得资金，路易斯·费利佩的夏季项目将使他能够在 将差异基因/蛋白质表达整合到HIS分段管道中，从而构建 个体发育过程中细胞命运谱系图和位置相关基因表达的检测。 路易斯·费利佩将获得实验程序和计算方法方面的专业知识，与 一名博士后在实验室进行实验室指导，并定期与PI会面。他还将参加 提高他的项目管理、科学沟通和团队合作能力。《副刊》 将使Luis Felipe能够完成他的项目并生成第一作者论文所需的数据，使他 竞争生物医学研究领域的顶级博士项目。如果他以博士生身份加入拉特克利夫实验室， 该项目将使他在博士论文研究方面领先一步，并使他在以下建议中更具竞争力 会为他的博士研究提供资金。

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.