Patent ID: 11920131
Assignee: TORAY INDUSTRIES, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method of producing a hairpin single-stranded RNA molecule capable of inhibiting expression of a target gene, the method comprising:
an annealing step of annealing a first single-stranded oligoRNA molecule and a second single-stranded oligoRNA molecule; and
a ligation step of ligating 3′ end of the first single-stranded oligoRNA molecule and 5′ end of the second single-stranded oligoRNA molecule by an Rnl2 family ligase,
wherein the first single-stranded oligoRNA molecule comprises a first RNA region and a second RNA region connected via a first linker, and one of the first RNA region and the second RNA region is capable of complementarily binding to the other,
the second single-stranded oligoRNA molecule comprises a third RNA region and a fourth RNA region connected via a second linker, and one of the third RNA region and the fourth RNA region is capable of complementarily binding to the other,
the first linker and the second linker are non-nucleotide linkers,
the first single-stranded oligoRNA molecule and the second single-stranded oligoRNA molecule are capable of forming an intermolecular double strand between complementary sequences at 5′-end or 3′-end thereof,
when the double strand is formed between the first single-stranded oligoRNA molecule and the second single-stranded oligoRNA molecule during the annealing step, a nick occurs between the 3′-end ribonucleotide residue of the first single-stranded oligoRNA molecule and the 5′-end ribonucleotide residue of the second single-stranded oligoRNA molecule, and a gap of at least one ribonucleotide residue is present between the 5′-end ribonucleotide residue of the first single-stranded oligoRNA molecule and the 3′-end ribonucleotide residue of the second single-stranded oligoRNA molecule, and
a sequence produced by ligating the first single-stranded oligoRNA molecule and the second single-stranded oligoRNA molecule comprises a gene expression-inhibiting sequence for the target gene.