Patent ID: 8512945

Claim:
A method of cleaving an exogenous non-tRNA target RNA molecule in vitro comprising the steps of: (a) providing a first RNA molecule within a mammalian cell, the first RNA molecule being the exogenous non-tRNA target RNA molecule; (b) contacting the first RNA molecule with a second RNA molecule, wherein the second RNA molecule comprises a targeting region consisting of: (i) a core region that is 7 nucleotides in length, the core region consisting of a 4-nucleotide sequence that is complementary to a 4-nucleotide sequence on the first RNA molecule, and a 3-nucleotide sequence that is directly 3′ or 5′ to the 4-nucleotide sequence, wherein the 3-nucleotide sequence is not complementary to the 3-nucleotide sequence that is directly 5′ or 3′ to the complementary 4-nucleotide sequence on the first RNA molecule; (ii) a 5′ flanking region that is directly 5′ to the core region and a 3′ flanking region that is directly 3′ to the core region, wherein each flanking region consists of a nucleotide sequence that is complementary to a nucleotide sequence on the first RNA molecule that is directly 3′ or 5′ to the 4-nucleotide sequence of (i), with the proviso that for the flanking region that is on the end of the core region having the 4-nucleotide sequence, the flanking region is complementary to the nucleotide sequence on the first RNA molecule beginning with the fourth nucleotide from the complementary 4-nucleotide sequence, whereby the first RNA molecule and the second RNA molecule form an RNA complex comprising a 3-nucleotide unpaired bulge-4-nucleotide paired helix-3-nucleotide unpaired bulge conformation, wherein one bulge has a guanine/adenine dinucleotide and the other bulge has a uracil/adenine dinucleotide; and (c) exposing the RNA complex to a heterologous archaeal Methanococcus jannaschii tRNA splicing endonuclease, wherein the target RNA molecule is cleaved.