Engineering a genetic construct to express soluble proteins tethered to the cell membrane

Abstract

Abstract Synthetic biology has significantly grown and developed in the last decades, allowing its implementation in a wide variety of fields and applications. However, there is a lack of versatile, modular tools that allow the expression of soluble proteins anchored to the cell membrane, which could be useful in a range of applications. Therefore, using a combination of computational simulations and experimental validation, we designed a genetic construct for the expression of soluble proteins tethered to the cell membrane. Using green fluorescent protein as a model soluble protein, we demonstrated that it was successfully tethered to the cell membrane and that it could be incorporated into polymer lipid nanodiscs and purified through the histidine tag. Replacement of the reporter gene with another gene of interest could lead to its application in a wide variety of fields, from creating artificial signalling or enzymatic cascades to studying lipid and protein dynamics in the cell membrane.