Modulation of cell membrane functionalization with aggregates of oligodeoxynucleotides containing alkyl chain-modified uridines.

In this study, we prepared oligodeoxynucleotides (ODNs) containing the uridine base modified by an alkyl chain at the 5-position (AU) and characterized their aggregate formation, localization, and functions in cells. These experiments revealed that aggregates of these ODNs were readily transported into cells, but their localization was dependent upon the number of hydrophobic units. ODNs with one modified AU were transported in the cytosol, while ODNs with multiple AU modifications resulted in their accumulation at the cell membrane. We also examined the ability of the AU-modified ODNs to capture small molecules at the cell membrane and their cellular uptake. We positioned a thioflavin-T (ThT)-binding aptamer on the cell membrane by means of hybridization with ODNs with three AUs at the strand end. Treatment with ThT resulted in its efficient uptake into cells, due to the capture of the ThT by the aptamers on the cell membrane. Thus, we demonstrated the functionalization of cell membranes with modified ODNs and the efficient delivery of small molecules into the cells.

Hybridizing Oligonucleotides with Hydrophobic Peptide Nucleic Acids Assists Their Cellular Uptake through Aggregate Formation.

We applied hybridization between hydrophobic peptide nucleic acids (PNAs) and oligodeoxynucleotides (ODNs) to achieve their cellular uptake without any need for transfection reagents. We employed a pyrenyl unit as a hydrophobic functional group and introduced it at the terminus of the PNA strand. The pyrene-tethered PNA (PyPNA) strongly bound with its complementary ODNs to generate amphiphiles; the resulting hybrids formed aggregates that showed efficient cellular uptake and high biological stability. Aggregates containing a functional DNA aptamer that bound to the PyPNA penetrated the cell membrane smoothly, with the aptamer exerting its original function in living cells. Thus, PyPNA efficiently assisted the additive-free cellular uptake of ODNs.