Compositions and methods for the expression of nucleic acids
OwnerDavid P. Davis, Daniel C. Gray +1inventors
InventorsDavid P. Davis, Daniel C. Gray, Zhenyu Gu
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Abstract
Compositions and methods are provided herein for the expression of nucleic acids. Compositions and methods are also provided herein for inducible expression of nucleic acids in transgenic cells and animals using transposon-based nucleic acid constructs. Compositions and methods are also provided herein for modulation of endogenous gene expression.
Claims34
A nucleic acid construct comprising:
- (a) a first transcription unit comprising a polynucleotide operably linked to an inducible promoter, wherein the inducible promoter comprises one or more TetO sequences;
- (b) a second transcription unit comprising a coding sequence encoding a TetR;
- and
- (c) a pair of inverted repeats, wherein one of the inverted repeats is 5′ of (a) and
- (b), and the other of the inverted repeats is 3′ of (a) and (b).
15-38. (canceled)
A method of expressing a polynucleotide in a transgenic mammal, the method comprising:
- (a) introducing into a mammalian, non-human embryonic cell a nucleic acid construct comprising:
- (i) a first transcription unit comprising the polynucleotide operably linked to an inducible promoter, wherein the polynucleotide encodes a regulatory RNA specific for an endogenous gene;
- and
- (ii) a pair of piggyBac inverted repeats, wherein one of the inverted repeats is 5′ of (i), and the other of the inverted repeats is 3′ of (i);
- (b) introducing into the mammalian, non-human embryonic cell a coding sequence encoding a piggyBac transposase that acts on the inverted repeats to mediate nucleic acid transposition;
- (c) generating a transgenic mammal from the mammalian, non-human embryonic cell into which the nucleic acid construct and the coding sequence encoding the transposase have been introduced;
- and
- (d) administering to the transgenic mammal an inducing agent that induces expression of the polynucleotide from the inducible promoter.
40-47. (canceled)
A transgenic non-human mammal comprising a nucleic acid construct comprising:
- (a) a first transcription unit comprising a polynucleotide operably linked to an inducible promoter, wherein the polynucleotide encodes a regulatory RNA specific for an endogenous gene;
- and
- (b) a pair of piggyBac inverted repeats, wherein one of the inverted repeats is 5′ of (a), and the other of the inverted repeats is 3′ of (a),
- wherein the regulatory RNA inhibits expression of the endogenous gene in the transgenic non-human mammal.
(canceled)
Dependencies are inferred from the claim text.