Gene regulation in transgenic animals using a transposon-based vector
View Patent ↗Administration of modified transposon-based vectors has been used to achieve stable incorporation of exogenous genes into animals. These transgenic animals produce transgenic progeny. Further, these transgenic animals produce large quantities of desired molecules encoded by the transgene. Transgenic egg-laying animals produce large quantities of desired molecules encoded by the transgene and deposit these molecules in the egg.
1. A vector comprising a nucleic acid sequence as set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 42, or SEQ ID NO: 43, wherein each sequence comprises:
(a) a prokaryotic transposase gene operably linked to a first promoter, wherein the nucleic acid sequence 3′ to the first promoter comprises the Kozak sequence as set forth in SEQ ID NO: 13, the Kozak sequence being positioned so as to include at least the first codon of the transposase gene, wherein the transposase gene is modified such that a plurality of the codons of the transposase gene that encode for amino acids 2-10 of a transposase protein encoded by the transposase gene are individually modified from the wild-type sequence of cytosine or guanine at the third base position of the codon to an adenine or a thymine, such that the modification does not change the amino acid encoded by the modified codon, and wherein the first promoter is a viral or a eukaryotic promoter;
(b) one or more genes of interest operably-linked to one or more additional promoters, wherein at least one of the genes of interest encodes for proinsulin or human growth hormone; and
(c) insertion sequences recognized by a transposase encoded by the modified transposase gene, wherein the transposon insertion sequences are positioned to flank the one or more genes of interest and their operably-linked promoters.
2. A vector comprising the nucleic acid sequence as set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 41, SEQ ID NO: 42, or SEQ ID NO: 43.