Development of a Cell Surface Display System inChlamydomonas reinhardtii

Abstract

Cell-surface display systems are biotechnological techniques used to expose heterologous proteins on the cell surface. Their application depends directly on the cell system used for the development, as well as anchoring point surface displayed protein. To meet most application demands an inexpensive, safe, and scalable production platform, that reduces the economic barriers for large scale use. Towards this goal, we screened three possible cell surface anchoring points in the green algae Chlamydomonas by fusing mVenus to prospective anchors moieties. The vectors harboring mVenus:anchor were screened for mVenus fluorescence and tested for cellular localization by confocal laser scanning microscopy. This strategy allowed the identification of two functional anchors, one for the cytoplasmic membrane using the MAW8 GPI-anchor signal, and one for the cell wall using the GP1 protein. We also exploited GP1 chemical and biological traits to release the fused proteins efficiently during cell wall shedding. Our work provides the base for surface engineering of C reinhardtii supporting both cell biology studies and biotechnology applications.