Enhancement of in vitro translation by nanoparticle conjugates
Provided herein are kits and methods suitable for enhancing in vitro translation of a nuclei acid sequence of interest.
1. A method for in vitro translation of an mRNA sequence of interest comprising contacting nanoparticles with an mRNA sequence of interest to be translated, and with reagents to cause in vitro translation, wherein the nanoparticles are conjugated to a nucleobase sequence selected from the group consisting of an RNA sequence, a DNA sequence and a PNA sequence, wherein the nucleobase sequence is complementary to at least a portion of the mRNA sequence, thereby resulting in translation of the mRNA of interest.
2. The method of claim 1 , wherein the nanoparticles are first contacted with the reagents and incubated prior to addition of the mRNA sequence of interest.
3. The method of claim 1 , wherein the nanoparticles are first contacted with the mRNA sequence of interest and incubated prior to addition of the reagents.
4. The method of claim 1 , wherein the nanoparticles are contacted with the mRNA sequence of interest at a ratio of less than one nanoparticle per mRNA sequence.
5. A method for in vitro translation of an mRNA sequence of interest comprising contacting nanoparticles with the mRNA sequence of interest to be translated, and with reagents to cause in vitro translation, wherein the nanoparticles are coated with PEG chains, thereby resulting in translation of the mRNA of interest.
6. The method of claim 5 , wherein each nanoparticle is coated with about 10 to 200 PEG chains.
7. The method of claim 5 , wherein the nanoparticles are further conjugated to a nucleobase sequence selected from the group consisting of an RNA sequence, a DNA sequence, and a PNA sequence, wherein the nucleobase sequence is complementary to at least a portion of the mRNA sequence.
8. The method of claim 1 or 5 , wherein the nanoparticles comprise gold.
9. The method of claim 1 or 5 , wherein the nanoparticles have a mean diameter of 5-25 nm.
10. The method of claim 1 or 5 , wherein the nanoparticles have a mean diameter of 10 nm.
11. The method of claim 1 or 7 , wherein the nucleobase sequence comprises a DNA sequence.
12. The method of claim 1 or 7 , wherein the nucleobase sequence comprises an RNA sequence.
13. The method of claim 1 or 7 , wherein the nucleobase sequence comprises a PNA sequence.
14. The method of claim 1 or 7 , wherein the nucleobase sequence is complementary to a non-translated portion of the mRNA sequence.
15. The method of claim 1 or 7 , wherein the nucleobase sequence is complementary to the Kozak Sequence.
16. The method of claim 1 or 7 , wherein the nucleobase sequence comprises a poly T sequence.
17. The method of claim 1 or 7 , wherein the nucleobase sequence is 10 to 50 bases in length.
18. The method of claim 1 or 7 , wherein each nanoparticle is conjugated to 10 to 100 nucleobase sequence strands.
19. The method of claim 1 or 7 , wherein each nanoparticle is conjugated to 20 to 70 nucleobase sequence strands.