Luciferase plasmid

The following procedure works well for the synthetic system developed by Sharp and colleagues (Doench et al., 2003). It uses a Renilla luciferase plasmid (based on pRL-TK, Promega, WI) harboring 4 concatenated, partially mismatched target sites for the artificial CXCR4 miR in its 3 UTR (p-RL-TK-4 sites). 

Figure 12.7 Nuclear import sequences increase gene expression in murine muscle. Mouse tibialis muscles were injected with 10 pg of CMV-driven luciferase plasmids either lacking (open bars) or containing (shaded bars) the SV40 enhancer downstream of the luciferase gene. Immediately after injection, the muscles were electroporated (8 pulses, 10 milliseconds each, 200V/cm) and the animals were allowed to recover. Three or 14 days later, muscles were removed and assayed for luciferase expression (average SEM,n=5).

Figure 11.17 Ternary phase diagram of complex coacervation between pRE-luciferase plasmid and chitosan at 55°C in 50 mmol dm No2S04. Sodium sulfate solution was regarded as one component, since the concentration change in the experiment range was minimal. The region to the right of line the ABC depicts the conditions under which phase separation occurs. The concentration ranges in the small grid area yield distinct particles.

While the applications of SELP-mediated controlled gene delivery are numerous, we have focused our efforts on controlled delivery to solid tumors. Our initial studies in this arena involved the intratumoral delivery of a reporter plasmid Renilla luciferase) to solid tumors in a murine (athymic nu/nu) model of human breast cancer (MDA-MB-435 cell line) (50). Delivery of the Renilla luciferase plasmid from SELP-47K matrices resulted in significantly enhanced tumor transfection for up to 21 days compared to naked DNA (Fig. 5). In particular, delivery of the plasmid from matrices containing 4 or 8 wt.% polymer resulted in enhanced transfection... 

Scaffolds can also be used for the delivery of genes. Kido et al. prepared scaffolds of gelatin and (3-tricalcium phosphate ((3-TCP) to search their gene transfection capabilities. The scaffolds were loaded with a spermine-pullulan luciferase plasmid DNA polyion complex and then stem cells were seeded into the scaffolds. It was shown that the level of plasmid DNA transfection was dependent on the method of the scaffold preparation. ... 

Fig. 9.26 Relative efficiency of generations 1-5 of dendrimers 13-G and 23-G compared to linear PEI for transfection of 3T3 cells with the luciferase plasmid cells viability with 13-G ...

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