Retrotransposon-mediated Gene Transfer for Animal Cells

Gene delivery methods for animal cells are one of the most important tools in biotechnology fields such as pharmaceutical protein production, generation transgenic animals and gene therapy. Because retrotransposons can move their own sequences to new genomic locations by a “copy-and-paste” process known retrotransposition, we attempted develop novel transfer system based on retrotransposon. A full-length long interspersed element-1 (LINE-1) contains 5’ untranslated region (5’UTR), two non-overlapping open reading frames (ORFs) separated short inter-ORF sequence, 3’UTR terminating an adenosine-rich tract. We constructed LINE-1 vector plasmid including components necessary retrotransposition. An intron-disrupted Neo reporter scFv-Fc expression unit under control CMV promoter were added into order evaluate retrotransposition express scFv-Fc. CHO-K1 transfected with plasmids screened G418. The established cell clones produced proteins culture medium. To steadily, also retrotransposon systems that supply ORF2 or ORF1–2 separately. Genomic PCR analysis revealed transgene derived from positive all clones. All tested