Nucleic acid functionalized nanoparticles for therapeutic applications
Materials and methods for modulating cellular uptake of functionalized nanoparticles are provided. Also provided are materials and methods for modulating the effectiveness of a therapeutic agent with a functionalized nanoparticle.
1. A method of increasing cellular uptake of a first plurality of nanoparticles relative to a second plurality of nanoparticles, the method comprising:
producing the first plurality of nanoparticles comprising a first mixed monolayer on the surface of the nanoparticles, the first mixed monolayer comprising a first oligonucleotide and a first diluent, wherein the first plurality consists essentially of nanoparticles having oligonucleotide functionalized on the nanoparticle at surface density X which is at least about 1.5 picomoles/cm 2 (pmol/cm 2 ) to about 25 pmol/cm 2 ;
producing the second plurality of nanoparticles comprising a second mixed monolayer on the surface of the nanoparticles, the second mixed monolayer comprising a second oligonucleotide and a second diluent, wherein the second plurality consists essentially of nanoparticles having oligonucleotide functionalized on the nanoparticle at surface density Y which is at least about 0.3 pmol/cm 2 to about 20 pmol/cm 2 ;
wherein X is greater than Y;
contacting a first cell with the first plurality and contacting a second cell with the second plurality;
wherein uptake of the first plurality of nanoparticles by the first cell is increased relative to uptake of an equal concentration of the second plurality by a second cell, wherein the first cell and the second cell are of the same type, wherein the first oligonucleotide and the second oligonucleotide do not comprise a cell-specific recognition element, and wherein the nanoparticles in the first plurality of nanoparticles and the nanoparticles in the second plurality of nanoparticles have a mean diameter of from about 5 to about 50 nanometers.
2. The method of claim 1 wherein the nanoparticles in the first plurality and the nanoparticles in the second plurality are each non-metallic.
3. The method of claim 1 wherein the nanoparticles in the first plurality and the nanoparticles in the second plurality are each metallic.
4. The method of claim 3 wherein the nanoparticles in the first plurality and the nanoparticles in the second plurality are each a colloidal metal.
5. The method of claim 4 wherein the nanoparticles in the first plurality and the nanoparticles in the second plurality are each gold.
6. The method of claim 1 wherein X is at least about 3 picomoles per centimeter 2 (pmol/cm 2 ).
7. The method of claim 1 wherein X is at least about 6 picomoles per centimeter 2 (pmol/cm 2 ).
8. The method of claim 1 wherein X is at least about 10 picomoles per centimeter 2 (pmol/cm 2 ).
9. The method of claim 1 wherein X is about 25 picomoles per centimeter 2 (pmol/cm 2 ).
10. The method of claim 1 wherein the first and or the second cell is a human cell.
11. The method of claim 1 wherein the first and/or the second diluent is oligo ethylene glycol (OEG).
12. The method of claim 1 wherein uptake of the first plurality is at least 5% greater than uptake of the second plurality.
13. The method of claim 1 wherein the first and/or the second monolayer comprises a less than 10-fold molar excess of diluent relative to oligonucleotide.
14. The method of claim 1 wherein the first and/or the second monolayer comprises a less than 20-fold molar excess of diluent relative to oligonucleotide.
15. The method of claim 1 wherein the first and/or the second monolayer comprises a less than 30-fold molar excess of diluent relative to oligonucleotide.
16. The method of claim 1 wherein the first and/or the second monolayer comprises a less than 40-fold molar excess of diluent relative to oligonucleotide.
17. The method of claim 1 wherein the first and/or the second monolayer comprises a less than 50-fold molar excess of diluent relative to oligonucleotide.
18. The method of claim 1 wherein the first and/or the second oligonucleotide is small interfering RNA (siRNA).