Two-Photon 3D Laser Printing Inside Synthetic Cells [Research Data]

Towards the ambitious goal of manufacturing synthetic cells from the bottom up, various cellular components have already been reconstituted inside of lipid vesicles. However, the deterministic positioning of these components inside the compartment has remained elusive. Here, by using two-photon 3D laser printing, 2D and 3D hydrogel architectures were manufactured with high precision and nearly arbitrary shape inside of preformed giant unilamellar lipid vesicles (GUVs). The required water-soluble photoresist is brought into the GUVs by diffusion in a single mixing step. Crucially, femtosecond two-photon printing inside the compartment does not destroy the GUVs. Beyond this proof-of-principle demonstration, early functional architectures were realized. In particular, a transmembrane structure acting as a pore was 3D printed, thereby allowing for the transport of biological cargo, including DNA, into the synthetic compartment. These experiments show that two-photon 3D laser microprinting can be an important addition to the existing toolbox of synthetic biology.