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. 2018 Dec 27;47(4):e23. doi: 10.1093/nar/gky1286

Figure 1.

Figure 1.

pMVP enables rapid, high-fidelity assembly of custom multicomponent transgene vectors. (A) The pMVP platform provides an array of Entry plasmids (pENTR) for promoters (6 options), 5′ modifiers (27 options), and 3′ modifiers (65 options) that can be partnered with a gene of interest and rapidly recombined into (B) an assortment of cross-compatible destination vectors (35 options) via an overnight recombination reaction to form (C) a customized experimental vector. In total, >108,000 unique vector permutations are possible for a gene of interest that is incorporated into pMVP. The resulting adenovirus (Ad), lentivirus (Lenti), Expression plasmid, Sleeping Beauty Transposon (SB), and PiggyBac transposon (PB) experimental vectors can be utilized in a broad variety of experimental contexts. (D) pMVP-derived vectors containing an EF1a promoter driving eGFP-P2A-PDX1-3xHA and either a polyA or WPRE were generated and added to primary (rat islet) or immortalized (HEK293) cells for either transient (adenovirus; expression) or stable (lentivirus, Sleeping Beauty, PiggyBac) transgene expression experiments. Cellular lysates were harvested for immunoblot analysis at the indicated times after viral transduction or plasmid transfection.