IP Library Granted Patent US 9,388,445
Granted Patent B2
US 9,388,445 · App. 13/868,648 · Granted Jul 12, 2016

Enhancement of in vitro translation by nanoparticle conjugates

Inventors: Kimberly S. Hamad-Schifferli (Somerville, MA); Sunho Park (Silver Spring, MD)
Assignee: Massachusetts Institute of Technology
C12P21/00C12P21/02B82Y5/00
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Quick Facts
Patent No.
US 9,388,445
App. No.
13/868,648
Granted
Jul 12, 2016
Kind
B2
Abstract

Provided herein are kits and methods suitable for enhancing in vitro translation of a nuclei acid sequence of interest.

Claims (32)

1. A kit for in vitro translation of an mRNA sequence of interest comprising:

a) nanoparticles, 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, and

b) one or more in vitro translation reagents for conducting in vitro translation of the mRNA sequence of interest, wherein at least one of the in vitro translation reagents is a lysate.

2. The kit of claim 1 , wherein the nanoparticles comprise gold.

3. The kit of claim 1 , wherein the nanoparticles have a mean diameter of 5-25 nm.

4. The kit of claim 1 , wherein the nanoparticles have a mean diameter of 10 nm.

5. The kit of claim 1 , wherein the nucleobase sequence comprises an RNA sequence.

6. The kit of claim 1 , wherein the nucleobase sequence is complementary to a non-translated portion of the mRNA sequence.

7. The kit of claim 1 , wherein the nucleobase sequence is complementary to the Kozak Sequence.

8. The kit of claim 1 , wherein the nucleobase sequence comprises a poly T sequence.

9. The kit of claim 1 , wherein the nucleobase sequence comprises a DNA sequence.

10. The kit of claim 1 , wherein the nucleobase sequence is 10 to 50 bases in length.

11. The kit of claim 1 , wherein each nanoparticle is conjugated to 10 to 100 nucleobase sequence strands.

12. The kit of claim 1 , wherein each nanoparticle is conjugated to 20 to 70 nucleobase sequence strands.

13. A kit for in vitro translation of sequence of interest comprising:

a) nanoparticles, wherein the nanoparticles are coated with polyethylene glycol (PEG) chains; and

b) one or more in vitro translation reagents for conducting in vitro translation of the mRNA of interest, wherein at least one of the in vitro translation reagents is a lysate.

14. The kit of claim 13 , wherein each nanoparticle is coated with 10 to 200 PEG chains.

15. The kit of claim 13 , 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.

16. The kit of claim 1 , wherein the nucleobase sequence comprise a PNA sequence.

17. The kit of claim 13 , wherein the nanoparticles comprise gold.

18. The kit of claim 13 , wherein the nanoparticles have a mean diameter of 5-25 nm.

19. The kit of claim 13 , wherein the nanoparticles have a mean diameter of 10 nm.

20. The kit of claim 15 , wherein the nucleobase sequence comprises an RNA sequence.

21. The kit of claim 15 , wherein the nucleobase sequence comprises a DNA sequence.

22. The kit of claim 15 , wherein the nucleobase sequence comprises a PNA sequence.

23. The kit of claim 15 , wherein the nucleobase sequence is complementary to a non-translated portion of the mRNA sequence.

24. The kit of claim 15 , wherein the nucleobase sequence is complementary to the Kozak Sequence.

25. The kit of claim 15 , wherein the nucleobase sequence comprises a poly T sequence.

26. The kit of claim 15 , wherein the nucleobase sequence is 10 to 50 bases in length.

27. The kit of claim 15 , wherein each nanoparticle is conjugated to 10 to 100 nucleobase sequence strands.

28. The kit of claim 15 , wherein each nanoparticle is conjugated to 20 to 70 nucleobase sequence strands.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 1, 2015
From: HAMAD-SCHIFFERLI, KIMBERLY S.; PARK, SUNHO
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 036702/0297 →
CONFIRMATORY LICENSE Recorded May 3, 2013
From: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 030342/0346 →
Continuity (3)
Continuation 12637743 · Dec 14, 2009
Provisional Application 61261733 · Nov 16, 2009
Related Publication 20140120575A1 · May 1, 2014