Double-stranded RNA stabilized in planta
First Claim
1. A recombinant DNA construct for plant cell transformation, comprising transcribable DNA that transcribes to a single RNA for silencing one or more target genes of a pest or pathogen of a plant, said DNA comprising operably linked two or more non-identical pairs of self-complementary sequences of said target genes,wherein said RNA comprises double-stranded RNA and has a stabilizing feature comprising multiple double-stranded RNA stems, said multiple double-stranded RNA stems comprising two or more non-identical pairs of self-complementary sequences, said stabilizing feature imparting improved resistance to a plant Dicer or Dicer-like enzyme relative to an RNA lacking said stabilizing feature, andwherein said one or more target genes of a pest or pathogen of a plant comprises a vacuolar ATPase gene, wherein the construct comprises SEQ ID NO:
- 16 or SEQ ID NO;
17,wherein said pest or pathogen of a plant is a Diabrotica species.
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Accused Products
Abstract
This invention discloses recombinant DNA constructs that transcribe to RNA having improved resistance to a plant RNase III enzyme, and methods for their use in making transgenic plant cells, plants, seeds, and products derived therefrom. Also provided are compositions and methods for imparting to a plant resistance to a pest or pathogen of the plant.
32 Citations
29 Claims
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1. A recombinant DNA construct for plant cell transformation, comprising transcribable DNA that transcribes to a single RNA for silencing one or more target genes of a pest or pathogen of a plant, said DNA comprising operably linked two or more non-identical pairs of self-complementary sequences of said target genes,
wherein said RNA comprises double-stranded RNA and has a stabilizing feature comprising multiple double-stranded RNA stems, said multiple double-stranded RNA stems comprising two or more non-identical pairs of self-complementary sequences, said stabilizing feature imparting improved resistance to a plant Dicer or Dicer-like enzyme relative to an RNA lacking said stabilizing feature, and wherein said one or more target genes of a pest or pathogen of a plant comprises a vacuolar ATPase gene, wherein the construct comprises SEQ ID NO: - 16 or SEQ ID NO;
17,wherein said pest or pathogen of a plant is a Diabrotica species. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 29)
- 16 or SEQ ID NO;
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25. A recombinant DNA construct comprising operably linked multiple anti-sense DNA segments that are anti-sense to multiple target sequences of a vacuolar ATPase gene and multiple sense DNA segments that are the same as said multiple target sequences, wherein when said recombinant DNA construct is transcribed to a single RNA, said multiple anti-sense DNA segments and the multiple sense DNA segments are transcribed to form multiple double-stranded RNA stems comprising two or more non-identical pairs of self-complementary sequences of said vacuolar ATPase gene wherein the construct comprises SEQ ID NO:
- 16 or SEQ ID NO;
17. - View Dependent Claims (26, 27, 28)
- 16 or SEQ ID NO;
Specification