Polynucleotides for causing RNA interference and method for inhibiting gene expression using the same
First Claim
1. A polynucleotide for causing RNA interference against a target gene selected from the genes of a target organism, which has at least a double-stranded region, wherein one strand in the double-stranded region consists of a base sequence homologous to a prescribed sequence which is contained in the base sequences of the target gene and which conforms to the following rules (a) to (d):
- (a) The 3′
end base is adenine, thymine or uracil;
(b) The 5′
end base is guanine or cytosine;
(c) A 7-base sequence from the 3′
end is rich in one or more types of bases selected from the group consisting of adenine, thymine and uracil; and
(d) The number of bases is within a range that allows RNA interference to occur without causing cytotoxicity, andwherein the other strand in the double-stranded region consists of a base sequence having a sequence complementary to the base sequence homologous to the prescribed sequence.
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Abstract
The present invention provides a polynucleotide that not only has a high RNA interference effect on its target gene, but also has a very small risk of causing RNA interference against a gene unrelated to the target gene. A sequence segment conforming to the following rules (a) to (d) is searched from the base sequences of a target gene for RNA interference and, based on the search results, a polynucleotide capable of causing RNAi is designed, synthesized, etc.:
- (a) The 3′ end base is adenine, thymine, or uracil,
- (b) The 5′ end base is guanine or cytosine,
- (c) A 7-base sequence from the 3′ end is rich in one or more types of bases selected from the group consisting of adenine, thymine, and uracil, and
- (d) The number of bases is within a range that allows RNA interference to occur without causing cytotoxicity.
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Citations
32 Claims
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1. A polynucleotide for causing RNA interference against a target gene selected from the genes of a target organism, which has at least a double-stranded region, wherein one strand in the double-stranded region consists of a base sequence homologous to a prescribed sequence which is contained in the base sequences of the target gene and which conforms to the following rules (a) to (d):
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(a) The 3′
end base is adenine, thymine or uracil;(b) The 5′
end base is guanine or cytosine;(c) A 7-base sequence from the 3′
end is rich in one or more types of bases selected from the group consisting of adenine, thymine and uracil; and(d) The number of bases is within a range that allows RNA interference to occur without causing cytotoxicity, and wherein the other strand in the double-stranded region consists of a base sequence having a sequence complementary to the base sequence homologous to the prescribed sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
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13. A method for selecting a polynucleotide to be introduced into an expression system for a target gene whose expression is to be inhibited, wherein the polynucleotide has at least a double-stranded region, wherein one strand in the double-stranded region consists of a base sequence homologous to a prescribed sequence which is contained in the base sequences of the target gene and which conforms to the following rules (a) to (f):
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(a) The 3′
end base is adenine, thymine or uracil;(b) The 5′
end base is guanine or cytosine;(c) A 7-base sequence from the 3′
end is rich in one or more types of bases selected from the group consisting of adenine, thymine and uracil;(d) The number of bases is within a range that allows RNA interference to occur without causing cytotoxicity; (e) A sequence in which 10 or more bases of guanine or cytosine are continuously present is not contained; and (f) A sequence sharing at least 90% homology with the prescribed sequence is not contained in the base sequences of genes other than the target gene among all gene sequences of the target organism, and wherein the other strand in the double-stranded region consists of a base sequence having a sequence complementary to the base sequence homologous to the prescribed sequence. - View Dependent Claims (14, 16)
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Specification