Phase II IUCRC at UF: Center for Arthropod Management Technologies - Single Cell Sequencing of Midguts from Pest Insects

ABSTRACT Recent advances in RNA sequencing at the single-cell level allow profiling of gene expression of individual cells to identify different cell types in complex tissues such as the midgut. The midgut is a complex tissue composed of multiple cell types performing many functions such as digestion, absorption of nutrients, production of hormones, and defense against microorganisms. Recent studies in Drosophila melanogaster adult intestine identified 22 distinct clusters of cells, including stem cells, enteroblasts, enteroendocrine cells, and enterocytes (1). We will use these recent developments in the field to perform scRNA-seq of larval or adult midgut from lepidopteran and hemipteran pest insects, respectively. The fall armyworm (FAW) and southern green stink bug (SGSB) or any other pest insects chosen by IAB could be used as model insects for Lepidoptera and Hemiptera. The main objectives of this project are 1. Optimization of methods for dissociation and isolation of single cells, preparation of libraries, and pipeline for analysis of sequence data using lepidopteran midgut as a model system. 2. Apply methods developed to perform single-cell sequencing of the hemipteran midgut. 3. Study cellular targets and mechanisms of Bt action by comparing scRNA-seq from Bt treated and untreated insects.