| Project Detail |
Reducing animal use with advanced mouse gene editing Mouse models have long been essential in life sciences research. However, these models present challenges, including difficulties with cell-type specific manipulations. There are also high maintenance costs and ethical concerns. Issues such as leaky promoters, off-target effects, and strain variability further complicate results. There is an urgent need for alternative strategies that can overcome these limitations. The ERC-funded LIMITLESS project aims to revolutionise mouse model research by developing a technology for whole-embryo gene editing in utero. Building on previous advances in neural plate targeting, the project will combine this with next-generation lineage tracing to map all cell types in developing embryos. This approach promises to reduce animal usage and enable experiments previously unattainable. Mouse models are a gold standard for life sciences research, spanning developmental biology, cancer studies, and pre-clinical testing, but they also have important limitations. Mouse colony maintenance is expensive, and the large colonies needed are ethically problematic. Approaches aiming for cell-type specific manipulations can be confounded by leaky promoters, off-target/unknown promoter expression, endogenous Cre sites, and Cre toxicity. Finally, mouse strains generated in different inbred genetic backgrounds yield confounding results when interbred, or necessitate extensive back-crossing, and again - large usage of mice. An alternative strategy to generate mouse models, circumventing these issues, is direly needed. To solve this problem for the mouse nervous system, we previously developed neural plate targeting with in utero nano-injection (NEPTUNE). NEPTUNE targets 95% of cells in the brain, can recapitulate established mouse models, and can be used flexibly for cell-type specific manipulations. In LIMITLESS, my lab will leverage our expertise in ultra-high titer viral production, developmental biology, and technology development to go beyond the state of the art and develop a technology for whole-embryo editing in utero. We will combine this technology with next generation single cell lineage tracing, to resolve the “family tree” of all cell types in developing embryos, providing both fundamental new knowledge and information that is leverageable for further development of the technique. In sum, my lab will go develop a much-needed technology for whole embryo genetic manipulation, opening the door to limitless new types of experiments. Direct in utero whole-embryo gene editing would accelerate life sciences research, reduce the financial and ethical costs of animal work, and enable experiments currently beyond reach with existing technology. |
