Methods and compositions for RNA interference
AssigneeUniversity of Utah Research Foundation
InventorsA. Alvarado, Peter Reddien, Adam Bermange
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Abstract
The invention relates to improved methods of attenuating expression of a target gene in a eukaryotic cell with dsRNA, identifying nucleic acid sequences responsible for conferring a particular phenotype to a cell, alleviating pest infestation in plants, and altering gene expression in an undifferentiated stem cell or the differentiated progeny thereof. Transcription of the RNA, which will form the dsRNA, is terminated by one or more terminators sequences, thereby increasing the efficiency of inhibition.
Claims53
A method of attenuating expression of a target nucleotide sequence in a eukaryotic cell, said method comprising:
- introducing double-stranded RNA (dsRNA) into the eukaryotic cell to attenuate expression of the target nucleotide sequence;
- wherein the dsRNA comprises a nucleotide sequence that hybridizes under stringent conditions to the target nucleotide sequence;
- and
- wherein said dsRNA is expressed from a vector containing one or more transcription terminators.
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A dsRNA for inhibiting expression of a gene, said dsRNA comprising:
- a first nucleotide sequence that hybridizes under stringent conditions to a target sequence, wherein the target sequence is complementary to said first nucleotide sequence, and wherein said stringent conditions include a wash step of 0.2×SSC at 65° C.
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Dependencies are inferred from the claim text.