A Large-Scale venture to Reduce the use of mice to perform DNA-Protein interaction assays

How genes are regulated is becoming incresingly studied, as this is the key to understand cell pathophysiology, and for the identification of therapeutic molecular targets. Protagonists of gene regulation are proteins called transcription factors (TFs). TFs bind the DNA on regulatory regions across the genome, and turn genes ON and OFF.

Knowledge of the genome-wide DNA-binding behaviour of TFs is therefore needed to understand how the genome functions.However, the study of TFs during organ formation and disease cannot be always modeled in vitro in a reliable fashion; moreover, the underlying biochemical procedure for these experiments requires significant amounts of tissue from laboratory animals.

In a relevant example, our past “success” demanded the sacrifice of ca. 250 mouse embryos to profile a single TF.This generates an urgent need of protocols aimed at reducing the number of animals employed. In this project, we will develop improved procedures starting from a novel technology called CUT&RUN-LoV-U.

Our aim is to increase its sensitivity and reduce the material required. Our preliminary experiments allowed us to successfully profile the DNA-binding behavior of TFs using