Stem Cells International / 2011 / Article / Fig 1

Research Article

Efficient Non-Viral Integration and Stable Gene Expression in Multipotent Adult Progenitor Cells

Figure 1

Schematic diagram of the internally controlled pKT2/NAG vector and integration products mediated by Sleeping Beauty (SB) transposase and by bacteriophage PhiC31 ( 𝜙 C31) integrase. (a) The integrating vector consists of (i) a 𝜙 C31 integrase recognition site (attB); (ii) eF1α or CMV- (Pr-) regulated GFP expression unit; (iii) neomycin resistance gene (Neo) transcriptionally regulated by the PGK promoter; (iv) flanking T2 transposase binding sites (IR/DRs; boxes with double triangles) separated by; (v) a colE1 bacterial origin of replication and kanamycin resistance gene; (vi) pA, polyadenylation signal from the rabbit beta globin gene. (b) SB transposase-mediated integration (left): the SB transposase excises transposon sequences at IR/DR transposase binding sites and precisely inserts them into TA-dinucleotide targets in cellular chromosomes, which are subsequently duplicated. 𝜙 C31 integrase-mediated integration (right): exogenous gene sequences on an attB containing plasmid integrate into mammalian genomes at “pseudo-attP” sites, chromosomal sequences having partial homology to the wild-type phage attP sequence.

Article of the Year Award: Outstanding research contributions of 2020, as selected by our Chief Editors. Read the winning articles.