Human RNase III and compositions and uses thereof
AssigneeIsis Pharmaceuticals, Inc.
InventorsHongjiang Wu, Stanley T. Crooke
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Abstract
The present invention provides polynucleotides encoding human RNAse III and polypeptides encoded thereby. Methods of using said polynucleotides and polypeptides are also provided.
Claims52
An isolated human RNase polypeptide comprising human RNase III.
A composition which comprises a human RNase III polypeptide and a pharmaceutically acceptable carrier.
An isolated polynucleotide which encodes a human RNase III polypeptide.
A vector which comprises a nucleic acid which encodes a human RNase III polypeptide.
A composition which comprises a vector comprising a nucleic acid encoding a human RNase III polypeptide and a pharmaceutically acceptable carrier.
An antibody targeted to a human RNase III polypeptide.
A nucleic acid probe at least 12 nucleobases in length which is capable of hybridizing to a portion of a nucleic acid encoding a human RNase III polypeptide.
An antisense compound 8 to 50 nucleobases in length which is targeted to a nucleic acid molecule encoding human RNase III, wherein said antisense compound specifically inhibits the expression of human RNAse III.
A method of inhibiting human RNase III expression or activity in a cell or tissue comprising contacting said cell or tissue with an inhibitor of human RNase III expression or activity.
A method of identifying agents which increase or decrease activity or levels of a human RNase III polypeptide in a host cell comprising:
- (a) contacting a cell expressing a human RNase III polypeptide with an agent suspected of increasing or decreasing activity or levels of the human RNase III polypeptide;
- and
- (b) measuring the activity or levels of the human RNase III polypeptide in the presence and absence of the agent so that an increase or decrease in the activity or levels of the human RNase III polypeptide can be determined.
A method of promoting inhibition of expression of a selected protein by an antisense oligonucleotide targeted to a target RNA encoding the selected protein comprising:
- (a) providing an antisense oligonucleotide targeted to a target RNA encoding a selected protein whose expression is to be inhibited;
- (b) allowing said oligonucleotide and said target RNA to hybridize to form an oligonucleotide-target RNA duplex;
- and
- (c) contacting said oligonucleotide-target RNA duplex with an RNase III polypeptide, under conditions in which cleavage of the target RNA strand of the oligonucleotide-target RNA duplex by the RNase III polypeptide occurs,
- whereby inhibition of expression of the selected protein is promoted.
A method of screening oligonucleotides to identify effective antisense oligonucleotides for inhibition of expression of a selected target protein comprising:
- (a) contacting a human RNase III polypeptide with a target RNA encoding the selected target protein and an oligonucleotide complementary to at least a portion of the target RNA under conditions in which an oligonucleotide-target RNA duplex is formed;
- (b) detecting cleavage of the target RNA of the oligonucleotide-target RNA duplex wherein cleavage is indicative of antisense efficacy.
A method of prognosticating efficacy of antisense therapy of a selected disease comprising measuring the level or activity of a human RNase III in a target cell of the antisense therapy.
A method of identifying agents which increase or decrease activity or levels of a human RNase III polypeptide in a host cell comprising:
- (a) contacting a cell expressing a human RNase III polypeptide with an agent suspected of increasing or decreasing activity or levels of the human RNase III polypeptide;
- and
- (b) measuring the activity or levels of the human RNase III polypeptide in the presence and absence of the agent so that an increase or decrease in the activity or levels of the human RNase III polypeptide can be determined.
A method of eliciting cleavage of a selected cellular RNA target comprising:
- (a) providing an antisense oligonucleotide targeted to a selected cellular RNA target to be cleaved, wherein said oligonucleotide is an RNA-like oligonucleotide;
- (b) allowing said oligonucleotide and said RNA to hybridize to form an oligonucleotide-RNA duplex;
- and
- (c) contacting said oligonucleotide-RNA duplex with an RNase III polypeptide, under conditions in which cleavage of the oligonucleotide-RNA duplex by the RNase III polypeptide occurs,
- whereby cleavage of the cellular RNA target is elicited.
A method of promoting RNA interference in a cell comprising providing to said cell an RNase III polypeptide in amounts sufficient to promote RNA interference.
A method of promoting RNA interference in a cell comprising enriching the amount or activity of RNase III polypeptide in said cell to a level sufficient to promote RNA interference.
Dependencies are inferred from the claim text.