This three-year, $1 million Project Grant from the National Science Foundation's Biological Sciences program (CFDA 47.074) supports the development of viral vectors for gene delivery and manipulation in amphibian research. The Trustees of Columbia University will lead efforts to design new viral vectors with host specificity for amphibians. Researchers at Columbia, the Institute of Science and Technology Austria, the University of Utah, and the Scripps Research Institute will collaborate to test vector libraries in Xenopus and Pleurodeles tissues and brains. They aim to generate vectors that can cross the blood-brain barrier and infect neurons systemically. University of Utah will also contribute to knockout native gene experiments in Xenopus eyes using CRISPR constructs delivered via viral vectors. All awardees will verify knockouts using validated antibodies and share protocols to facilitate vector development in non-model organisms. This work seeks to provide new genetic and molecular tools to strengthen amphibian research.