Methods and materials for producing transgenic animals
AssigneeRecombinectics, Inc.
InventorsSCOTT C. FAHRENKRUG, DANIEL F. CARLSON, ARON M. GEURTS
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Abstract
Transgenic artiodactyls are described as well as methods of making and using such artiodactyls.
Claims42
A transgenic pig, the nucleated cells of which comprise a nucleic acid construct, said nucleic acid construct comprising a transcriptional unit comprising a regulatory region operably linked to a nucleic acid sequence encoding a small interfering RNA that reduces expression of porcine cystic fibrosis transmembrane conductance regulator (CFTR), wherein an inverted repeat of a transposon flanks each side of said transcriptional unit.
A method for making a transgenic pig, said method comprising:
- a) introducing a transgenic pig cell into an enucleated pig oocyte to establish a combined cell, wherein said transgenic pig cell comprises a nucleic acid construct and a source of a transposase, said nucleic acid construct comprising a transcriptional unit, said transcriptional unit comprising a regulatory region operably linked to a nucleic acid sequence encoding a small interfering RNA that reduces expression of porcine cystic fibrosis transmembrane conductance regulator (CFTR), wherein an inverted repeat of a transposon flanks each side of said transcriptional unit;
- b) producing a porcine embryo from said combined cell;
- c) transferring said porcine embryo to a recipient female;
- and
- d) allowing said porcine embryo to develop in the recipient female to produce said transgenic pig, the nucleated cells of which comprise said nucleic acid construct.
A method for making a transgenic artiodactyl, said method comprising:
- a) injecting a nucleic acid construct and a source of a transposase into a pronuclear staged in vitro fertilized egg, said nucleic acid construct comprising a transcriptional unit, said transcriptional unit comprising a regulatory region operably linked to a target nucleic acid sequence, wherein an inverted repeat of a transposon flanks each side of said transcriptional unit;
- b) transferring said injected fertilized egg to a recipient female;
- and
- c) allowing said injected fertilized egg to develop in said recipient animal female to produce said transgenic artiodactyl.
A method for making a transgenic artiodactyl, said method comprising:
- a) injecting at least two nucleic acid constructs and a source of a transposase into a pronuclear staged in vitro fertilized egg, wherein each said nucleic acid construct comprises a transcriptional unit and an inverted repeat of a transposon flanking each side of said transcriptional unit, wherein said transcriptional unit of one of said nucleic acid construct comprises a regulatory region operably linked to a target nucleic acid sequence, and wherein said transcriptional unit of one of said nucleic acid constructs comprises a regulatory region operably linked to a nucleic acid sequence encoding a selectable marker;
- b) transferring said injected fertilized egg to a recipient female;
- and
- c) allowing said injected fertilized egg to develop in said recipient female to produce said transgenic artiodactyl.
A transgenic artiodactyl, the nucleated cells of which comprise a nucleic acid construct, said nucleic acid construct comprising a transcriptional unit comprising a regulatory region operably linked to a nucleic acid sequence encoding a small interfering RNA that reduces expression of a target gene.
A nucleic acid construct comprising a hypermethylated transposon.
Dependencies are inferred from the claim text.