Patent ID: 9238805
Filing Date: 2016-01-19
CPC Classification: C12N,C12P,C12Y

Claim Text:
1. A method of sequence specific cleavage of dsRNA substrate by a dsRNA endoribonuclease, comprising the steps of a) combining the dsRNA endoribonuclease with dsRNA substrate in a mixture, wherein said dsRNA endoribonuclease comprises amino acid sequence of SEQ ID NO:1 with a D94R mutation; and has the loop that is locating in and interacting with a major groove of dsRNA, which corresponds to the loop locating in and interacting with a major groove of dsRNA in the model of structure of endoribonuclease Mini III in complex with dsRNA; and wherein said dsRNA endoribonuclease exhibits the dsRNA sequence specific activity within the consensus sequence (SEQ ID NOS: 111 and 121) 5′ DACCUHD 3′ 3′ HUGGADH 5′ where H=A or C or U; D=A or G or U; preferably said dsRNA endoribonuclease exhibits the dsRNA sequence specific activity within the consensus sequence SEQ ID NOS: 112 and 122) 5′ YGACCUCGNNG 3′ 3′ RCUGGAGCNNC 5′ where Y=C or U; R=A or G; N=G or A or U or C; and wherein the specific sequence in dsRNA substrate recognized by said dsRNA endoribonuclease is the consensus sequence (SEQ ID NOS: 111 and 121) 5′ DACCUHD 3′ 3′ HUGGADH 5′ where H=A or C or U; D=A or G or U; preferably is the consensus sequence (SEQ ID NOS: 112 and 122) 5′ YGACCUCGNNG 3′ 3′ RCUGGAGCNNC 5′ where Y=C or U; R=A or G; N=G or A or U or C, b) cleaving the said dsRNA substrate within said recognition sequence by said dsRNA endoribonuclease.