DNA aptamer targeting zinc transporters ZIP10 and ZIP6 on cancer cells

Cell-SELEX is an effective method for generating aptamers that specifically bind molecules in their native state on live cells. It not only uncovers novel potential biomarkers but also provides robust molecular recognition tools for a wide spectrum of applications. In this work, we generate a high affinity aptamer, HL15, through Cell-SELEX. A refined sequence HL15a demonstrated a strong binding affinity to target cells, with a minimal dissociation constant (K_d) of just 1.90 ± 0.49 nM. Subsequent truncation and mutation assays revealed that the core sequence of HL15a forms an antiparallel G-quadruplex structure. Furthermore, the target proteins of aptamer HL15a were identified and confirmed to be ZIP10 and ZIP6, both members of the zinc transporter ZIP family with high homology. Using HL15a as a molecular probe, we detected a range of universal binding affinities to the majority of tumor cells in the 48 cell lines evaluated. Furthermore, HL15a was effectively applied to distinguish high malignancy cancer tissues from both normal and low malignancy tissues in the pathological sections of breast and prostate cancers. Given that ZIP10 and ZIP6 play crucial roles in regulating zinc homeostasis and are implicated in numerous diseases, the aptamer HL15a offers a powerful tool for the study and potential therapeutic intervention of these two proteins.