No abstract

Spherical nucleic acids (SNAs) are spherically arranged oligonucleotides on core inorganic nanoparticles and have great potential for intracellular delivery of bioactive molecules, since they have been found to be internalized into mammalian cells. Understanding the factors that influence the cellular uptake of SNAs would be beneficial to design SNAs with novel uptake properties. We here report the effect of the sugar backbone type of the oligonucleotides on the cellular internalization of SNAs. After the preparation of SNAs with the oligonucleotides of five different sugar backbones, we analyze the cellular uptake efficiency quantitatively by flow cytometry and inductively coupled plasma mass spectrometry (ICP-MS). The data reveal that the uptake efficiencies and the uptake mechanisms significantly rely on the backbone type. These results suggest that the backbone modification can provide a unique handle to tune the cellular uptake behavior of SNAs.

The tetracycline repressor (TetR)-regulated system is a widely used tool to study gene functions through control of its expression. Various effectors such as tetracycline (Tc) and doxycycline (Dox) quickly induce or shut down gene expression, but reversing gene expression has not been eligible due to long half-lives of such effectors. Here, we found that procaspase activating compound 1 (PAC-1) rapidly reduces transient expression of TetR-regulated green fluorescent protein (GFP) in mammalian cells. Next, we applied PAC-1 to control of expression of transient receptor potential melastatin 7 (TRPM7) protein, whose downstream cellular events can be monitored by cell morphological changes. We observed that PAC-1 quickly reduces TRPM7 expression, consequently affecting cell morphology regulated by TRPM7. The present study demonstrates the first small molecule that efficiently turns off the TetR-regulated gene expression in mammalian cells, thereby precisely regulating the expression level of target gene.