Patent ID: 11898186
Assignee: GENSCRIPT USA INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC A  C

Claim 0:
1. A method for in vitro transcription of a DNA template into RNA, comprising
providing (1) a DNA template comprises a promoter operably linked to a nucleic acid comprising a 5′ untranslated region (5′ UTR), an open reading frame (ORF) encoding the RNA of interest, a 3′ UTR, and a poly A region, and
(2) a cap analogue comprises the structure of, wherein R1 and R2 are each CH3 or H; and B1 and B2 are each A, U, G, or C,
wherein the promoter comprises the sequence of
TAATACGACTCACTATAX1X2X3 (SEQ ID NO: 16),
wherein A at position 17 is −1 nucleotide and X1 at position 18 is +1 nucleotide,
when X1 is G, X2 and X3 are each A, T, G, or C, then B1 is A and B2 is G,
when X1 is C, X2 and X3 are each A, T, G, or C, then B1 is A and B2 is C, and
when X1 is T, X2 and X3 are each A, T, G, or C, then B1 is A and B2 is U,

wherein the cap analogue binds to −1 and +1 nucleotides of the promoter, and

incubating the DNA template and the cap analogue in a reaction mixture, wherein the incubating comprises incubating the reaction mixture at from about 15° C. to about 35° C. for from about 1 hour to about 12 hours, thereby producing the RNA, wherein
the 5′ UTR consists of SEQ ID NO: 1 and the 3′ UTR consists of SEQ ID NO: 4, or
the 5′ UTR consists of SEQ ID NO: 1 and the 3′ UTR consists of SEQ ID NO: 6, or
the 5′ UTR consists of SEQ ID NO: 3 and the 3′ UTR consists of SEQ ID NO: 4, or
the 5′ UTR consists of SEQ ID NO: 3 and the 3′ UTR consists of SEQ ID NO: 6, or
the 5′ UTR consists of SEQ ID NO: 3 and the 3′ UTR consists of SEQ ID NO: 8, or
the 5′ UTR consists of SEQ ID NO: 5 and the 3′ UTR consists of SEQ ID NO: 4, or
the 5′ UTR consists of SEQ ID NO: 5 and the 3′ UTR consists of SEQ ID NO: 6, or
the 5′ UTR consists of SEQ ID NO: 7 and the 3′ UTR consists of SEQ ID NO: 4, or
the 5′ UTR consists of SEQ ID NO: 7 and the 3′ UTR consists of SEQ ID NO: 6, or
the 5′ UTR consists of SEQ ID NO: 7 and the 3′ UTR consists of SEQ ID NO: 8, or
the 5′ UTR consists of SEQ ID NO: 9 and the 3′ UTR consists of SEQ ID NO: 4, or
the 5′ UTR consists of SEQ ID NO: 9 and the 3′ UTR consists of SEQ ID NO: 6.