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. Author manuscript; available in PMC: 2019 Mar 21.
Published in final edited form as: Semin Cell Dev Biol. 2018 Mar 31;86:140–149. doi: 10.1016/j.semcdb.2018.03.019

Fig. 7. Retrotransposition assay.

Fig. 7.

A) Organization of an L1 vector in a typical retrotransposition assay. The L1 vector contains an antisense copy of the neo gene disrupted by an intron in the sense orientation. sd and sa are the splice donor and splice acceptor sites, respectively. The intron of the transcribed L1 vector will be spliced and contain the antisense copy of the neo gene. Retrotransposition competent elements will support subsequent cDNA synthesis of this transcript at a DNA target site and ultimately the insertion of an active copy of the neo gene, which when expressed from its promoter (Pr, in red) generates colonies of G418 resistant cells or foci. B) An example of stained foci generated from the retrotransposition assay described in (A) [43]. p111_rtc, p151_rtc and p555_rtc are the L1 vectors containing the modern human, mosaic and the resuscitated primate ORF1 sequences (see inset of Fig. 6B), respectively.