Our Technology
Summary
Transposagen Biopharmaceuticals has pioneered the use of mobile DNA technology (e.g., the DNA transposons Sleeping Beauty and piggyBac) to generate insertional mutations in the rat germ line and is now in possession of several rat stem cell technologies that can be combined synergistically with mobile DNA technology, representing by far the most cost effective method to create knockout rat lines.
Rat Stem Cells
Transposagen is developing several inbred and outbred rat stem cell lines for the creation of TKO™ Knockout Rat Models. The company has exclusive rights to rat spermatagonial stem cells, which offer significant advantages over rat embryonic stem cells and rat induced pluripotent stem cells in terms of speed and efficiency that a knockout animal can be created.
For more information email info@transposagenbio.com or call 859-428-8561.
piggyBac
piggyBac (PB) DNA transposons mobilize via a "cut-and-paste" mechanism whereby a transposase enzyme, encoded by the transposon itself, excises and re-integrates the transposon at other sites within the genome. The amino acid PB transposase specifically recognizes PB inverted terminal repeats (ITRs) that flank the transposon; it binds to these sequences and catalyzes excision of the transposon. PB then integrates at TTAA sites throughout the genome, in an relatively random fashion. For the creation of gene trap mutations (or adapted for generating transgenic animals), the transposase is supplied in trans on one plasmid and is co-transfected with a plasmid containing donor transposon, a recombinant transposon comprising a gene trap flanked by the binding sites for the transposase (ITRs). The transposase will catalyze the excision of the transposon from the plasmid and subsequent integration into the genome. Integration within a coding region will capture the elements necessary for gene trap expression.
If you have an interest in sub-licensing piggyBac technology, please email ordering@transposagenbio.com or call 859-428-8561.
Sleeping Beauty
The sleeping beauty (SB) transposon is a derivative of the Tc1/mariner superfamily of DNA transposons prevalent among both vertebrate and invertebrate genomes. However, endogenous DNA transposons from this family are completely inactive in vertebrate genomes. An active Tc1/mariner transposon, synthesized from alignment of inactive transposons from the salmonid subfamily of elements, was “awoken” to form the transposon named Sleeping Beauty. SB, like other DNA transposons, mobilizes itself via a cut-and-paste mechanism whereby a transposase enzyme, encoded by the transposon itself, excises and re-integrates the transposon at other sites within the genome. The 340 amino acid SB protein recognizes inverted terminal repeats (ITRs) that flank the transposon; it binds to these sequences and catalyzes excision of the transposon. SB then integrates into random sites within the genome, although some studies report very slight preferences for transcriptional units. There is also a simple requirement of a TA-dinucleotide at the target site, like all Tc1/mariner transposons.
The SB transposon is a powerful tool for insertional mutagenesis in many vertebrate species. It recently exhibited especial utility for germ line mutagenesis in both mice and rats. There are several advantages that make SB a highly attractive mutagen geared toward gene discovery: 1) it has little bias for inserting within particular genomic regions or within specific recognition sequences, 2) de novo insertions of the transposon provide a “tagged” sequence marker for rapid identification of the specific mutation by simple PCR cloning methods, 3) in vivo SB insertional mutagenesis allows multiple mutations to be quickly and easily generated in a single animal, and in a single tissue, such as an adenomatous polyp.