Ribozyme for identifying modification on rna sequence and rna cleavage method using same

A hammerhead ribozyme-target RNA construct represented by the general formula [III]:

• (wherein A means adenine (A);
• C means cytosine (C);
• G means guanine (G);
• U means uracil (U);
• D is adenine (A), cytosine (C), guanine (G) or uracil (U), and means a base which forms a base pair with H;
• X is a modification recognition base for recognizing the modification site of target RNA, which means adenine (A) or cytosine (C), provided however that X means adenine (A) when E is cytosine (C) and X means cytosine (C) when E is adenine (A);
• N may be the same or different and is a base selected from adenine (A), cytosine (C), guanine (G) and uracil (U), which forms a base pair with N′;
• E is a base selected from adenine (A), cytosine (C), guanine (G), uracil (U) and inosine (I), which forms a base pair with the corresponding base X in the general formula [III] and, when forming a base pair with the base X, means a base whose bond with the base H′ adjacent to the 5′ side is cleaved;
• H′ means a base selected from adenine (A), cytosine (C), guanine (G) and uracil (U);
• N′ may be the same or different and is a base selected from adenine (A), cytosine (C), guanine (G) and uracil (U), which forms a base pair with a corresponding base N;
• a means an integer from 1 to 10, preferably an integer from 2 to 6, and more preferably an integer from 2 to 4;
• b means an integer from 1 to 6, and preferably an integer from 2 to 4;
• c means an integer from 1 to 4, and preferably an integer from 1 to 2;
• m means an integer from 2 to 50, preferably an integer from 5 to 30, and further preferably an integer from 5 to 20;
• and
• n means an integer from 2 to 50, preferably an integer from 5 to 30, and further preferably an integer from 5 to 20;
• and
• the arrow symbol means a cleavage site).

A method for the cleavage of a target RNA comprising:

• reacting a hammerhead ribozyme represented by the general formula [I]:
• (wherein A, C, G, U, D, X, N, N′, a, b and c have each the same meaning as above) with a target RNA represented by the general formula [II]:
• 5′-N′-N′m-N′-H-H′-E-N′-N′n-N′-3′ [II]
• (wherein, E, N′, H, H′, m and n have the same meaning as above) to give a hammerhead ribozyme-target RNA construct represented by the general formula [III]:
• (wherein A, C, G, U, D, E, N, N′, H, H′, X, a, b, c, m and n have the same meaning as above).