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US 6,218,185 B1Granted
Piggybac transposon-based genetic transformation system for insects
AssigneesThe United States of America as represented by the Secretary of Agriculture, University of Notre Dame, University of Florida
InventorsPaul D Shirk, Malcolm J. Fraser, Jr., Teresa A. Elick, Omaththage P. Perera
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Abstract
The present invention is directed to nucleic acid and amino acid sequences for transformation constructs containing piggyBac or tagalong transposable elements. These constructs allow for the precise excision and insertion of heterologous DNA into a host cell.
Claims28
A transformation system comprising:
- (a) a first DNA comprising a non-transposon heterologous DNA sequence inserted between a pair of inverted repeats of a piggyBac transposon, and
- (b) a second DNA encoding a transposase active on the pair of inverted repeats,
- wherein the second DNA is incapable of transposition caused by said transposon.
A method of introducing a heterologous DNA sequence into the genome of a cultured cell of interest comprising:
- (a) introducing into the cell a first nucleic acid comprising a non-transposon heterologous DNA sequence inserted between a pair of inverted repeats of a piggyBac transposon;
- and
- (b) providing the cell with a second nucleic acid that encodes a transposase active on the pair of inverted repeats such that the heterologous DNA sequence is inserted into the genome of the cell, said second nucleic acid characterized in that it is incapable of transposition by the transposase.
A cell comprising:
- (a) a first DNA comprising a non-transposon heterologous DNA sequence inserted between a pair of inverted repeats of a piggyBac transposon;
- and
- (b) a second DNA encoding a transposase active on a pair of inverted repeats, which second DNA is incapable of transposition caused by the transposase.
A cell comprising:
- (a) a first DNA comprising a non-transposon heterologous DNA sequence operably linked to an inducible promoter and inserted between a pair on inverted repeats of a pigyyBac transposon;
- and
- (b) a second DNA encoding a transposase active on a pair of inverted repeats.
Dependencies are inferred from the claim text.