Stefanie Maes, Louis Delhaye, Robin Cooreman, Eline Soetens, Annick Verhee, Delphine De Sutter, Tessa Van de Steene, Long T. Nguyen, Dominique Audenaert, Xavier Saelens, Sven Eyckerman
Studying protein-protein interactions, which are crucial for all cellular processes in both health and disease, offers valuable insights into underlying mechanisms and reveals promising therapeutic targets. While efforts have been made to vastly increase the scale of MAPPIT assays, covering the full protein interactome remains challenging, in particular when considering the availability of different assay configurations. In this study, we expanded our Golden Gate modular cloning platform to enable rapid, customizable construct assembly for MAPPIT assays. We demonstrate the strength of this expanded toolbox for multiple distinct protein-protein interactions, including a MAPPIT assay for six different intracellular VHHs targeting the antiviral effector protein MXA.