Gene silencing by systemic rna interference
AssigneePresident and Fellows of Harvard College
InventorsCraig Hunter, William Winston, Evan Feinberg
Life of this patent
Estimating the term…
Abstract
Nucleic acid and protein sequences relating to a gene required for systemic RNAi are disclosed. The SID-1 and 2 proteins are shown to be required for systemic RNAi. Nucleic acids, vectors, transformed cells, transgenic animals, polypeptides, and antibodies relating to the <i>sid-1</i><i>and</i>2 genes and proteins are disclosed. Also provided are methods for reducing the expression of a target gene in a cell, a population of cells, or an animal.
Claims91
An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of:
- (a) at least 10 consecutive nucleotides of SEQ ID NO:
- 1;
- (b) at least 12 consecutive nucleotides of SEQ ID NO:
- 1;
- (c) at least 14 consecutive nucleotides of SEQ ID NO:
- 1;
- (d) at least 16 consecutive nucleotides of SEQ ID NO:
- 1;
- (e) at least 18 consecutive nucleotides of SEQ ID NO:
- 1;
- and
- (f) a sequence complementary to any one of the sequences of (a)-(e).
An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of:
- (a) a sequence encoding a SID-1 protein;
- (b) a sequence encoding at least a transmembrane domain of a SID-1 protein;
- (c) a sequence encoding at least an extracellular domain of a SID-1 protein;
- and
- (d) a sequence complementary to any one of the sequences of (a)-(c).
An isolated polypeptide comprising an amino acid sequence of SEQ ID NO. 2.
A nucleic acid that hybridizes to at least a portion of SEQ ID NO. 1.
A vector, comprising a sid-1 nucleotide sequence operably linked to a regulatory region such that the sid-1 nucleotide sequence is expressed in a host.
An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of:
- (a) at least 10 consecutive nucleotides of SEQ ID NO:
- 5;
- (b) at least 12 consecutive nucleotides of SEQ ID NO:
- 5;
- (c) at least 14 consecutive nucleotides of SEQ ID NO:
- 5;
- (d) at least 16 consecutive nucleotides of SEQ ID NO:
- 5;
- (e) at least 18 consecutive nucleotides of SEQ ID NO:
- 5;
- and
- (f) a sequence complementary to any one of the sequences of (a)-(e).
A vector, comprising a sid-2 nucleotide sequence operably linked to a regulatory region such that the sid-2 nucleotide sequence is expressed in a host.
A method for modifying a genome of an animal comprising the insertion of a nucleic acid selected from the group consisting of a nucleotide sequence encoding at least a fragment of SID-1 protein, at least a transmembrane portion of a SID-1 protein, and at least an extracellular domain of a SID-1 protein.
A method for modifying a genome of an animal comprising the insertion of a nucleic acid selected from the group consisting of a nucleotide sequence encoding at least a fragment of SID-2 protein.
An isolated polypeptide comprising a amino acid sequence selected from the group consisting of:
- (a) a sequence encoding a SID-1 protein;
- (b) a sequence encoding at least a transmembrane domain of a SID-1 protein;
- (c) a sequence encoding at least an extracellular domain of a SID-1 protein;
- and
- (d) a sequence complementary to any one of the sequences of (a)-(c).
An antibody that specifically binds to an isolated polypeptide that comprises an immunogenic fragment of SEQ ID NO. 2.
A kit for detecting at least one epitope of a polypeptide comprising SEQ ID NO. 2 or an antigenic fragment thereof having
- an antibody that specifically binds to a fragment of SEQ ID NO. 2;
- and
- reagents necessary for facilitating binding and detection of SEQ ID NO. 2.
An isolated polypeptide comprising an amino acid sequence of SID-2.
An antibody that specifically binds to an isolated polypeptide that comprises an immunogenic fragment of SID-2.
A kit for detecting at least one epitope of a polypeptide comprising SID-2 or an antigenic fragment thereof having
- an antibody that specifically binds to a fragment of SID-2;
- and
- reagents necessary for facilitating binding and detection of SID-2.
A method of restoring or initiating systemic RNA interference in an organism, comprising the introduction of a nucleic acid sequence selected from the group consisting of sid-1 and sid-2 nucleotide sequence to said organism.
A method for reducing the expression of a target gene in a cell, comprising:
- introducing a nucleic acid vector having a sid-1 nucleotide sequence into said cell such that sid-1 is expressed in said cell;
- introducing a double-stranded RNA molecule having a nucleotide sequence complementary to said target gene;
- and
- detecting the expression level of protein encoded by said target gene.
A method for reducing the expression of a target gene in a cell, comprising:
- introducing a nucleic acid vector having a sid-2 nucleotide sequence into said cell such that sid-2 is expressed in said cell;
- introducing a double-stranded RNA molecule having a nucleotide sequence complementary to said target gene;
- and
- detecting the expression level of protein encoded by said target gene.
A method for expressing a sid nucleic acid in an embryonic stem cell, comprising:
- obtaining said embryonic stem cell;
- transfecting said embryonic stem cell with a sid nucleic acid;
- and
- expressing said sid nucleic acid within said embryonic stem cells.
Dependencies are inferred from the claim text.