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US 2019/0144489 A1Published application
Methods and compositions for nucleoside triphosphate and ribonucleic acid production
AssigneeGreenLight Biosciences, Inc.
InventorsDrew S. Cunningham, Daniel MacEachran, James Robbins Abshire, Himanshu Dhamankar, Ifeyinwa Iwuchukwu, Mehak Gupta, Matthew Eduardo Moura, Naveen Sudharsan, Nicholas Skizim, Rachit Jain, Karthikeyan Ramachandriya
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Abstract
Provided herein, in some embodiments, are methods and composition for the production of nucleoside triphosphates and ribonucleic acids.
Claims31
-26. (canceled)
A method for producing ribonucleic acid (RNA), comprising:
- (a) incubating in a reaction mixture (i) cellular RNA and (ii) a ribonuclease to produce 5′ nucleoside monophosphates (NMPs);
- (b) eliminating the ribonuclease;
- and
- (c) incubating in the reaction mixture, or in a second reaction mixture, the 5′ NMPs, polyphosphate kinase (PPK), polyphosphate, deoxyribonucleic acid (DNA) template encoding a RNA of interest, and RNA polymerase to produce the RNA of interest.
1 more element
A method for producing ribonucleic acid (RNA), comprising:
- (a) incubating in a reaction mixture (i) cellular RNA and (ii) a ribonuclease to produce 5′ nucleoside monophosphates (NMPs);
- (b) eliminating the ribonuclease;
- and
- (c) incubating in the reaction mixture, or in a second reaction mixture, the 5′ NMPs, NMP kinases, NDP kinase, polyphosphate kinase (PPK), polyphosphate, deoxyribonucleic acid (DNA) template encoding a RNA of interest, and RNA polymerase to produce the RNA of interest.
1 more element
A method for producing ribonucleic acid (RNA), comprising:
- (a) incubating in a reaction mixture (i) cellular RNA and (ii) Nuclease P1 to produce 5′ nucleoside monophosphates (NMPs);
- (b) eliminating the Nuclease P1;
- and
- (c) incubating in the reaction mixture, or in a second reaction mixture, the 5′ NMPs, NMP kinases, NDP kinase, a Class III PPK2 enzyme from Deinococcus geothermalis (SEQ ID NO:
- 1), polyphosphate, deoxyribonucleic acid (DNA) template encoding a RNA of interest, and RNA polymerase to produce the RNA of interest.
2 more elements
A method for producing nucleoside triphosphates (NTPs) comprising:
- (a) incubating in a reaction mixture cellular RNA and a ribonuclease to produce 5′ nucleoside monophosphates (NMPs);
- and
- (b) incubating in the reaction mixture, or in a second reaction mixture, the 5′ NMPs, polyphosphate kinase, and polyphosphate.
Dependencies are inferred from the claim text.