A method of regulating gene expression

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.

1 .- 30 . (canceled) 
     
     
         31 . A cell comprising a target gene and a DNA construct comprising a constitutive or inducible promoter functional in said cell operably linked to a nucleic acid comprising a nucleotide sequence encoding, as part of a longer encoded sequence, an miRNA precursor, said miRNA precursor comprising a stem loop structure and comprising in said stem of said stem loop structure a sequence complementary to a portion of an RNA transcript of said target gene, wherein said stem of said stem loop structure is about 19-45 base pairs long and the loop of said stem loop structure is about 4-25 nucleotides long. 
     
     
         32 . The cell of  claim 31 , wherein following induction of said promoter in said cell:
 (i) said nucleotide sequence is transcribed,   (ii) the resulting transcript of said nucleotide sequence is processed so that said miRNA precursor is excised from said transcript of said nucleotide sequence,   (iii) said miRNA precursor is processed so that a mature miRNA about 21 or 22 nucleotides in length is excised from said miRNA precursor, and   (iv) inhibition of expression of said gene is effected.   
     
     
         33 . The cell of  claim 31 , wherein said promoter is a polymerase II-based promoter. 
     
     
         34 . The cell of  claim 31 , wherein the cell is a plant cell. 
     
     
         35 . A plant comprising the plant cell of  claim 34 . 
     
     
         36 . The cell of  claim 31 , wherein the cell is a non-human animal cell. 
     
     
         37 . A non-human animal comprising the cell of  claim 36 .