Down-regulation of gene expression using artificial micrornas for silencing fatty acid biosynthetic genes
AssigneeE. I. du Pont Nemours and Company
InventorsHoward Glenn Damude, Brian McGonigle
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Abstract
Isolated nucleic acid fragments comprising precursor miRNAs, and artificial miRNAs and their use in down-regulating gene expression of fatty acid biosynthetic genes are described.
Claims18
An isolated nucleic acid sequence comprising a sequence encoding at least one artificial microRNA precursor, operably linked to at least one regulatory sequence, wherein said sequence encoding an artificial microRNA precursor is transcribed so that a transcript comprising said artificial miRNA precursor is produced, and wherein said transcript is processed so that a mature miRNA about 21 to 22 nts in length is excised from said artificial miRNA precursor, and expression of at least one plant fatty acid biosynthetic gene(s) selected from the group consisting of:
- fad2-1, fad2-2, fad3, fatB, sad, and fael is inhibited.
A method for reducing expression at least one plant fatty acid biosynthetic gene, said method comprising:
- (a) transforming a plant cell with at least one isolated nucleic acid sequence comprising a sequence encoding at least one artificial microRNA precursor, operably linked to at least one regulatory sequence;
- (b) said at least one sequence encoding an artificial microRNA precursor is transcribed so that a transcript comprising said at least one artificial miRNA precursor is produced;
- (c) said transcript is processed so that at least one mature miRNA about 21 to 22 nts in length is excised from said artificial miRNA precursor, and
- (d) expression of at least one plant fatty acid biosynthetic gene selected from the group consisting of:
- fad2-1, fad2-2, fad3, fatB, sad, and fael is reduced.
An isolated nucleic acid sequence comprising a sequence encoding two or more artificial microRNA precursors, operably linked to at least one regulatory sequence, wherein said two or more artificial microRNA precursor can be on the same or separate transcriptional units, further wherein said sequence encoding artificial microRNA precursors is transcribed so that a transcript comprising said artificial miRNA precursors is produced, and wherein said transcript is processed so that mature miRNAs about 21 to 22 nts in length are excised from said artificial miRNA precursors, and expression of two or more plant fatty acid biosynthetic gene(s) selected from the group consisting of:
- fad2-1, fad2-2, fad3, fatB, sad, and fael is inhibited.
A method for reducing expression two or more plant fatty acid biosynthetic gene, said method comprising:
- (a) transforming a plant cell with at least one isolated nucleic acid sequence comprising a sequence encoding two or more artificial microRNA precursors, wherein said two or more artificial microRNA precursors can be on the same or separate transcriptional units, operably linked to at least one regulatory sequence;
- (b) said at least one sequence encoding two or more artificial microRNA precursors is transcribed so that at least one transcript comprising said two or more artificial miRNA precursors is produced;
- (c) said at least one transcript is processed so that at least two or more mature miRNAs about 21 to 22 nts in length are excised from said artificial miRNA precursors, and
- (d) expression of two or more plant fatty acid biosynthetic genes selected from the group consisting of:
- fad2-1, fad2-2, fad3, fatB, sad, and fael are reduced.
An isolated nucleic acid sequence comprising a sequence encoding at least one artificial microRNA precursor, operably linked to at least one regulatory sequence, wherein said sequence encoding at least one artificial microRNA precursor is capable of forming a double-stranded RNA or hairpin, wherein the at least one amiRNA precursor comprises at least one modified miRNA precursor in which the miRNA sequence and its complementary sequence are replaced by at least one amiRNA sequence and at least one STAR sequence, wherein expression of at least one plant fatty acid biosynthetic gene(s) selected from the group consisting of:
- fad2-1, fad2-2, fad3, fatB, sad, and fael is inhibited.
An isolated nucleic acid sequence comprising a sequence encoding two or more artificial microRNA precursors, operably linked to at least one regulatory sequence, wherein said two or more artificial microRNA precursor can be on the same or separate transcriptional units, further wherein said sequences encoding two or more artificial microRNA precursors are capable of forming a double-stranded RNA or hairpin, wherein the two or more amiRNA precursors comprise a modified miRNA precursor(s) in which the miRNA sequence(s) and its complementary sequence(s) are replaced by two or more amiRNA sequences and two or more STAR sequences, and further wherein expression of at least two or more plant fatty acid biosynthetic genes selected from the group consisting of:
- fad2-1, fad2-2, fad3, fatB, sad, and fael, are inhibited.
Dependencies are inferred from the claim text.