Value Proposition
· Enhanced germline transmission - Tol2 transposon system provides superior integration efficiency compared to episomal DNA and alternative transposon systems
· Streamlined cloning workflow - Gateway cloning system enables rapid insertion of test sequences without traditional restriction/ligation steps
· Real-time visualization – GFP tag allow for dynamic monitoring of expression patterns throughout embryogenesis
· Cross-species compatibility - Can evaluate regulatory sequences from any vertebrate species, including human elements
· High-throughput capability - Suitable for systematic screening of multiple candidate sequences
Unmet Need
Scientists estimate that a significant portion of the human genome contains regulatory sequences that control when and where genes are turned on, but identifying which specific DNA sequences are actually functional remains extremely challenging. Computational tools can predict regulatory DNA sequences genome-wide, but most predictions fail experimental validation due to high false-positive rates. Current validation methods rely on expensive, slow mouse transgenesis or cell culture assays lacking developmental context, creating major bottlenecks for large-scale functional genomics studies. This limits identification of regulatory mutations contributing to human disease, despite their recognized importance in complex genetic disorders. Therefore, there is a critical need for a high-throughput, cost-effective platform to systematically validate regulatory sequence function and accelerate genomic discovery translation.
Technology Description
Researchers at Johns Hopkins have developed a Tol2 transposon-based vector system that enables rapid functional analysis of regulatory DNA sequences in zebrafish embryos. The vector contains enhanced green fluorescent protein under a minimal promoter, allowing researchers to clone candidate regulatory sequences and directly visualize their activity through tissue-specific reporter expression, where GFP expression is only activated when the regulatory sequence being tested functions as an enhancer to drive the minimal promoter. Functional regulatory elements drive tissue-specific or temporally regulated EGFP expression that can be visualized throughout embryogenesis, providing a rapid and quantitative readout of regulatory sequence activity.
Stage of Development
· The disclosed technology is validated and ready for licensing.
Data Availability
· n/a
Publication
Shannon Fisher et al. Conservation of RET Regulatory Function from Human to Zebrafish Without Sequence Similarity.Science312,276-279(2006).DOI:10.1126/science.1124070
