MRNA functionalization method
The present invention provides a functionalized mRNA including mRNA and double-stranded RNA including at least one RNA oligomer hybridized with mRNA. Functionalized mRNA is provided according to this configuration.
1. An mRNA vaccine, which comprises a double-stranded RNA consisting of an mRNA encoding an antigen, at least one first RNA oligomer hybridizing with the mRNA, and a second RNA oligomer hybridizing with the first RNA oligomer, wherein
the first RNA oligomer comprises:
(a) an RNA sequence comprising a first RNA sequence consisting of a sequence of 12 to 40 nucleotides complementary to the sequence of the mRNA, and a second RNA sequence consisting of a sequence of 10 to 200 nucleotides complementary to the sequence of the second RNA oligomer, in this order from the 5′-terminus thereof,
(b) an RNA sequence comprising a first RNA sequence having an identity of 90% or more to a sequence of 12 to 40 nucleotides complementary to the sequence of the mRNA and hybridizing with the mRNA, and a second RNA sequence having an identity of 90% or more to a sequence of 10 to 200 nucleotides complementary to the sequence of the second RNA oligomer and hybridizing with the second RNA oligomer, in this order from the 5′-terminus thereof,
(c) an RNA sequence comprising a second RNA sequence consisting of a sequence of 10 to 200 nucleotides complementary to the second RNA oligomer, and a first RNA sequence consisting of a sequence of 12 to 40 nucleotides complementary to the sequence of the mRNA, in this order from the 5′-terminus thereof, or
(d) an RNA sequence comprising a second RNA sequence having an identity of 90% or more to a sequence of 10 to 200 nucleotides complementary to the sequence of the second RNA oligomer and hybridizing with the second RNA oligomer, and a first RNA sequence having an identity of 90% or more to a sequence of 12 to 40 nucleotides complementary to the sequence of the mRNA and hybridizing with the mRNA, in this order from the 5′-terminus thereof.
2. The mRNA vaccine according to claim 1 , wherein the first RNA oligomer consists of a sequence of 22 to 240 nucleotides.
3. The mRNA vaccine according to claim 1 , wherein the number of the first RNA oligomers to be allowed to hybridize with one mRNA is 1 to 50.
4. The mRNA vaccine according to claim 1 , wherein the first RNA oligomer comprises the RNA sequence (a) or the RNA sequence (b), and the second RNA oligomer has a triphosphoric acid structure at the 5′-terminus thereof.
5. The mRNA vaccine according to claim 1 , wherein the first RNA oligomer comprises the RNA sequence (c) or the RNA sequence (d), and the first RNA oligomer has a triphosphoric acid structure at the 5′-terminus thereof.
6. The mRNA vaccine according to claim 1 , wherein the terminus of the double-stranded RNA on the side at which the second RNA oligomer hybridizes with the first RNA oligomer is a blunt end.
7. The mRNA vaccine according to claim 1 , wherein the second RNA oligomer comprises a sequence of 10 to 200 nucleotides.
8. The mRNA vaccine according to claim 1 , wherein the double-stranded RNA is in a naked form.
9. The mRNA vaccine according to claim 1 , which is not used together with an adjuvant.
10. The mRNA vaccine according to claim 1 , which is for use in the prevention or treatment of a disease in a subject in need thereof.