Preparation And Isolation of 5' Capped mRNA
AssigneeLife Technologies Corporation
InventorsAnilkumar KORE, Shanmugasundaram MUTHIAN, Irudaya CHARLES
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Abstract
The synthesis of capped/tagged RNA, methods of use and kits providing same are contemplated. Tagged RNA permits isolation of RNA transcripts in vitro. The ability to isolate and purify capped RNA results in improved transcription and translation and provides a tool for identifying RNA-protein interactions. Such capped RNA finds use in therapeutic applications, diagnosis and prognosis and in the treatment of cancers and HIV.
Claims25
A composition comprising:
- wherein,
- B is a nucleobase;
- R1 is selected from a halogen, OH, and OCH3;
- R2 is selected from H, OH, and OCH3;
- R3 is CH3 or void;
- R4 is NH2;
- R5 is H;
- and
- n is 1, 2 or 3;
- and
- wherein,
- a linker is attached to one of R1, R2, R4, R5, or B.
A method of synthesizing a dinucleotide cap analog comprising:
- a) providing a guanosine nucleoside comprising either a 2′ substituent or a 3′ substituent and optionally comprising a linker;
- b) phosphorylating the nucleoside, forming a nucleotide;
- c) methylating the nucleotide;
- d) adding a phosphorylated second nucleotide optionally comprising a linker;
- e) coupling said first nucleotide with said second nucleotide, forming a dinucleotide cap analog.
A kit for capping an RNA transcript comprising a cap analog having the structure:
- wherein,
- B is a nucleobase;
- R1 is selected from a halogen, OH, and OCH3;
- R2 is selected from OH, OCH3, and H;
- R3 is CH3 or void;
- R4 is NH2;
- R5 is H;
- and
- n is 1, 2 or 3;
- and wherein,
- a linker is attached to one of R1, R2, R4, R5, or B;
- a reporter moiety is attached to the linker;
- at least R1, or R2 is OH;
- and
- b) an RNA polymerase.
Dependencies are inferred from the claim text.