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. 2016 Oct 4;5(10):e369. doi: 10.1038/mtna.2016.76

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Copyright © 2016 Official journal of the American Society of Gene & Cell Therapy

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Schematic of double fluorescent (DB) minimal piggyBac vector constructs: 1.6 kb (DB) (a) and 1.6 kb (DB-2) (b). Both plasmids contained minimal terminal repeats (5′TRmin, 3′TRmin), mCherry under control of the cytomegalovirus (CMV) promoter and a bGH polyadenylation signal (pA) in the delivered fragment (marked by vertical bars). The transposase fragment of the vectors contains the piggyBac transposase open reading frame (ORF) (PBase) with truncated terminal domains (5′TD (trunc.), 3′TD (trunc.)). PiggyBac transposase expression was controlled by the phosphoglycerate kinase (PGK) promoter as described above. Both plasmids had an additional transcription unit (not present in the previously described red fluorescent protein (RFP)-expressing vectors) that contained the simian virus 40 (SV40) promoter outside of the transposon and transposase fragment to drive green fluorescent protein (eGFP) expression. Plasmid 1.6 kb (DB-2) was identical to plasmid 1.6 kb (DB), but the SV40 promoter was replaced by the CMV promoter to drive eGFP. The expression of eGFP in both vectors was terminated by a SV40 polyadenylation signal (pA).