Lipid Anchor Chemistry and Spacer Architecture Influence DNA Availability and Light-Directed Cell Printing

Precise control over cellular interactions is critical to understanding the complex underpinnings of disease. DNA-printing of cells (DPC), a technology that uses complementary DNA to program cellular interactions, has the potential to enable precision cell printing. However, the simplicity of printing is undermined by the stability of anchoring moieties. This manuscript systematically investigates how commercially available hydrophobic DNA anchor and co-anchor chemistries influence DNA availability and light-directed DPC (LD-DPC) printability. Using cholesterol, tocopherol, and cholesterol-tocopherol modifications with defined spacer architectures, we evaluated how anchor chemistry and molecular architecture influence functional DNA presentation at the cell surface. Notably, incorporating additional polyethylene glycol spacers between lipid anchors and DNA enhances functional DNA availability and patterning performance. While cholesterol-based anchors provide the highest initial surface loading, tocopherol-containing anchors promote greater spatial dispersion and cell-capture efficiency during printing. Notably, multiple anchor/co-anchor combinations maintain functional DNA presentation at room temperature and in serum-containing media. These findings demonstrate that lipid anchor identity and spacer architecture strongly influence the availability of membrane-associated DNA and the performance of light-directed DNA printing of cells. Collectively, these results provide practical guidance for designing stable nucleic-acid-modified cell interfaces using commercially available lipid-DNA chemistries.