Targeting DNA
nanotiles to glycocalyx-anchored ssDNA initiators.
(a) Diagram showing the design and decoration of the DNA nanotile
with ssDNA-comp overhangs and biotin tags. (b) AFM images of assembled
DNA nanotiles without decoration and with full decorations (28 ssDNA-comp
overhangs and 35 biotin tags). (c) DNA gel electrophoresis analysis
of 2-Log DNA ladder, M13 scaffold, undecorated nanotiles, and fully
decorated nanotiles. (d) Azide ligands were metabolically incorporated
into glycans within the glycocalyx by administering Ac4ManNAz. These
cell-surface azide ligands were conjugated with azide-reactive 5′DBCO-ssDNA
initiators, leading to covalent immobilization of ssDNA initiators
onto the glycocalyx, which can then recruit DNA nanotiles via hybridization
with the complementary ssDNA-comp overhangs on nanotiles. (e–g)
ssDNA initiators, immobilized on the glycocalyx, were detected through
its hybridization with the fluorescent, complementary 5′FAM-ssDNA-comp
oligos (e). FAM fluorescence intensity was visualized using microscopic
imaging (f) and quantified using a spectrometer (g). (h, i) DNA nanotiles
with 35 biotin tags and 1, 3, 6, 14, or 28 complementary ssDNA-comp
overhangs were targeted to glycocalyx-anchored ssDNA initiators. Cell-surface
nanotiles were visualized via biotin detection using a fluorophore-conjugated
streptavidin (red) (h), and the fluorescence intensity was quantified
using a spectrometer (i). Data represent means ± s.d. from three
independent replicates. **P ≤ 0.01, ***P ≤ 0.001.