Collaborative Research: EDGE FGT: Cell type-specific tools for the study of gene function in sponges.

University Of Denver, Denver CO

Investigators

Abstract

Sponges are aquatic invertebrates with many important characteristics. Studying these animals provides insight into the evolution of complexity, specialized cell types, and interactions with microbes. Sponges also produce many natural compounds relevant to the pharmaceutical and biotechnology sectors. Yet despite their significance across these many scientific areas, research progress has been limited by the lack of tools to manipulate sponge genes and cells. This project builds on a recent technical advance that enables efficient delivery and expression of genetic material in sponges. This project will (1) develop a comprehensive library of molecular tools for visualizing and controlling gene activity in specific sponge cells, (2) establish standardized, reproducible protocols for genome editing, and (3) provide direct training opportunities to speed the adoption of these methods by researchers. All methods and tools will be made openly available to accelerate community-wide uptake. By transforming sponges into a system where genes can be readily studied, this project will empower new lines of investigation across developmental biology, immunology, cell evolution, and natural product discovery. At the core of this effort is the development of cell type-specific genetic tools for the freshwater sponge Spongilla lacustris, a widely used laboratory model in sponge biology. Building on a published single-cell atlas and a high-efficiency electroporation method, the research team will (1) optimize transfection conditions using a factorial design and quantitative reporter assays, (2) create a modular plasmid toolkit featuring cell type-specific promoters, codon-optimized fluorescent proteins, and subcellular targeting signals, and (3) implement CRISPR/Cas9 genome editing to generate stable transgenic lines. These tools will enable researchers to directly test gene function in vivo and investigate cellular mechanisms that have been inaccessible in sponges until now. The project will also serve as a model for extending genetic technologies to other experimentally challenging but scientifically important organisms. This research will develop new biotechnology that will be made available to the scientific community. 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.