Method of regulating gene expression
AssigneeDuke University
InventorsBryan R. Cullen, Yan Zeng
Life of this patent
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Abstract
The present invention relates, in general, to gene expression and, in particular, to a method of inhibiting the expression of a target gene and to constructs suitable for use in such a method.
Claims12
A plant cell comprising a DNA construct comprising a constitutive or inducible promoter operably linked to a nucleic acid comprising a sequence encoding an artificial micro RNA (miRNA) precursor, wherein said artificial miRNA precursor is a naturally occurring miRNA precursor, wherein a portion of the stem of the stem loop structure of said naturally occurring miRNA precursor is substituted by an exogenous sequence complementary to a portion of an RNA transcript of, a target gene in said cell,
- wherein, following introduction of said construct into said cell:
- i) said nucleic acid is transcribed so that a transcript comprising said artificial miRNA precursor is produced,
- ii) said transcript resulting from step (i) is processed so that said artificial miRNA precursor is excised therefrom,
- iii) said artificial miRNA precursor resulting from step (ii) is processed so that a mature miRNA about 21 or 22 nucleotides in length is excised from said artificial miRNA precursor, and
- iv) inhibition of expression of said gene is effected.
A non-human animal comprising, in at least one cell thereof, a DNA construct comprising a constitutive or inducible promoter operably linked to a nucleic acid comprising a sequence encoding an artificial micro RNA (miRNA) precursor, wherein said miRNA precursor is a naturally occurring miRNA precursor, wherein a portion of the stem of the stem loop structure of said naturally occurring miRNA precursor is substituted by an exogenous sequence complementary to a portion of an RNA transcript of a target gene in said cell,
- wherein, following introduction of said construct into said cell:
- i) said nucleic acid is transcribed so that a transcript comprising said artificial miRNA precursor is produced,
- ii) said transcript resulting from step (i) is processed so that said artificial miRNA precursor is excised therefrom,
- iii) said artificial miRNA precursor resulting from step is processed so that a mature miRNA about 21 or 22 nucleotides in length is excised from said artificial miRNA precursor, and
- iv) inhibition of expression of said gene is effected.
Dependencies are inferred from the claim text.