Enhanced selective expression of transgenes in fiber producing plants
AssigneesBay Cropscience AG, Texas Tech University
InventorsFrank Meulewaeter, Zhixin Xie, Gengxiang Jia, Arnab Ghosh, Forrest Sheng Bao
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Abstract
Methods and means are provided to enhance the selective expression of transgenes under control of a fiber-selective promoter, in fiber cells, particularly cotton fiber cells by including target sites for naturally occurring microRNAs with a specific expression profile, particularly with a differential expression profile between cells leading to fibers and other cells of the fiber producing plant, into the transcribed region of genes of interest.
Claims41
A recombinant gene for spatially selective expression in a cotton plant comprising the following operably linked elements:
- (a) a plant-expressible promoter;
- (b) a region encoding a biologically active RNA molecule;
- (c) optionally a 3’ transcription termination and polyadenylation region characterized in that said recombinant gene further comprises a heterologous target sequence recognized by an endogenous miRNA, said miRNA being differentially expressed in cells leading to fibers in said cotton plant compared to cells of said cotton plants other than said cells leading to said fibers.
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A target site having the nucleotide sequence of any one of SEQ ID Nos 5 to 8, SEQ ID No. 5 from nucleotide position 4 to nucleotide position 24, SEQ ID No. 6 from nucleotide position 4 to nucleotide position 24, SEQ ID No. 8, SEQ ID No. 8 from nucleotide position 4 to 24, SEQ ID No. 23, SEQ ID No. 23 from nucleotide position 4 to 24, SEQ ID No. 23 from nucleotide position 11 to 32, or SEQ ID No. 23 from nucleotide positions 4 to 32 or a nucleotide sequence complementary to a miRNA having a nucleotide sequence according to any one of SEQ ID Nos 1 to 4, SEQ ID No. 20, SEQ ID No. 21, SEQ ID No. 22 or SEQ ID No. 23 whereby one or more of the following mismatches may occur:
- (a) a mismatch between the nucleotide at the 5′ end of said miRNA and the corresponding nucleotide sequence in the target RNA sequence;
- (b) a mismatch between any one of the nucleotides in position 1 to position 9 of said miRNA and the corresponding nucleotide sequence in the target RNA sequence;
- (c) three mismatches between any one of the nucleotides in position 12 to position 21 of said miRNA and the corresponding nucleotide sequence in the target RNA sequence provided that there are no more than two consecutive mismatches;
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- (d) no mismatch is allowed at positions 10 and 11 of the miRNA.
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A pre-microRNA from cotton having a nucleotide sequence of any one of SEQ ID NOs:
- 9, 10, 11 or 12, or a nucleotide sequence essentially similar or identical thereto wherein the microRNA region and optionally the region complementary to the microRNA region (microRNA* region) have been altered.
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Dependencies are inferred from the claim text.