Down-regulation of gene expression using artificial micrornas
AssigneeE. I. Du Pont de Nemours and Company
InventorBrian McGonigle
Life of this patent
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Abstract
Isolated nucleic acid fragments comprising precursor miRNA, and artificial miRNAs and their use in down-regulating gene expression are described.
Claims7
An isolated nucleic acid fragment comprising the deoxyribonucleotide sequence set forth in SEQ ID NO:13 wherein (i) nucleotides 53 to 73 of SEQ ID NO:13 are replaced by a first variable nucleotide subsequence ranging in size from 19 to 24 nucleotides depending upon the target sequence whose expression is to be reduced, (ii) nucleotides 97 to 117 of SEQ ID NO:13 are replaced by a second variable nucleotide subsequence ranging in size from 19 to 24 nucleotides, said second variable nucleotide subsequence being capable of hybridizing to the first variable subsequence, and (iii) the precursor miRNA produced by said isolated nucleic acid fragment has the same stem structure as the precursor miRNA produced by endogenous SEQ ID NO:13.
A method for reducing expression of a target sequence in a plant cell, said method comprising:
- (a) transforming at least one plant cell with a nucleic acid construct comprising the deoxyribonucleotide sequence set forth in SEQ ID NO:13 wherein (i) wherein nucleotides 53 to 73 of SEQ ID NO:13 are replaced by a first variable nucleotide subsequence ranging in size from 19 to 24 nucleotides depending upon the target sequence whose expression is to be reduced, (ii) nucleotides 97 to 117 of SEQ ID NO:13 are replaced by a second variable nucleotide subsequence ranging in size from 19 to 24 nucleotides, said second variable nucleotide subsequence being capable of hybridizing to the first variable subsequence, and (iii) the precursor miRNA produced by said isolated nucleic acid fragment has the same stem structure as the precursor miRNA produced by endogenous SEQ ID NO:13;
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- (b) selecting those transformed plant cell(s) whose level of expression of the target sequence is reduced when compared to the level of expression of the target gene in a wild type plant cell.
Dependencies are inferred from the claim text.