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US 6,933,146 B2Granted
Methods and means for producing efficient silencing construct using recombinational cloning
AssigneeCommonwealth Scientific and Industrial Research
InventorsChristopher A. Helliwell, Susan V. Wesley, Peter M. Waterhouse
Life of this patent
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Abstract
Methods and vectors and kits are provided for producing chimeric nucleic acid constructs capable of producing dsRNA for silencing target nucleic acid sequences of interest using recombinational cloning.
Claims28
A vector comprising the following operably linked DNA fragments:
- an origin of replication allowing replication in a recipient cell (1),
- a selectable marker region (2) capable of being expressed in said recipient cell;
- and
- a chimeric DNA construct comprising in sequence:
- a promoter or promoter region (3) capable of being recognized by RNA polymerases of a eukaryotic cell;
- a first recombination site (4), a second recombination site (5), a third recombination site (6) and a fourth recombination site (7);
- a 3′ transcription terminating and polyadenylation region (8) functional in said eukaryotic cell:
- wherein
- said first recombination site (4) and said fourth recombination site (7) are capable of reacting with a same recombination site;
- and
- said second recombination site (5) and said third recombination site (6), are capable of reacting with a same recombination site;
- and wherein
- said first recombination site (4) and said second recombination site (5) do not recombine with each other or with a same recombination site;
- or
- said third recombination site (6) and said fourth recombination site (7) do not recombine with each other or with a same recombination site.
12 more elements
A vector comprising the sequence of SEQ ID No 13.
A vector comprising the sequence of SEQ ID No 23.
A vector comprising the sequence of SEQ ID No 24.
A vector comprising the sequence of SEQ ID No 25.
A vector comprising the sequence of SEQ ID No 26.
Dependencies are inferred from the claim text.