Vectors luciferase

FIGURE 20.2 The involvement of RARE on apo-lO -lycopenoic acid-transactivated RARp expression. Upper panel Diagram of the RARp reporter vector with wild type and mutated R AREs. Lower panel HeLa cells transfected with the RARp reporter vector and an internal control vector were treated with 5 pmol/I. of apo-lO -lycopenoic acid or 1 pmol/L of all-trans retinoic acid for 24h. Luciferase activities were measured by dual-luciferase reporter system. Values are means of SEM of three replicate assays., statistically significantly different, as compared with control in the same group, P < 0.05. (Adapted from Lian, F. et al., Carcinogenesis, 28, 1567, 2007. With permission.)... 

Luciferase reporter gene (pGL3 -control vector) activity measured in transfected fibroblast 183... 

Fig. 3. (A) SK-N-SH neuronal cells were cotransfected with Bcl-xL or Bcl-xL AkB luciferase reporter plasmids together with pSG-c-Rel and pSG-RelA, or a combination ofpSG-p50, pSG-RelA, andpSG-c-Rel expression plasmids. Control cells were transfected with empty pSG5 vector. Twenty-four hours later the luciferase activity was measured. Mutation of NF- B binding site blocked the luciferase expression. The c-Rel-containing dimers, p50/c-Rel and RelA/c-Rel, but not p50/RelA complex, were able to activate Bcl-xL promoter. Values are mean S.E.M. of three experiments run in triplicate ( /> < 0.05 vs. the corresponding control value). (B) OGD-induced NF-acB activation promotes Bim but not Bcl-xL transcription. Primary cortical neurons were transfected with Bim or Bcl-xL luciferase reporter plasmid or with Bim and Bcl-xL AkB luciferase reporter plasmids and then exposed to OGD. Four hours later, luciferase activity was measured. The OGD exposure significantly induced Bim and decreased Bcl-xL promoter activity. Mutation of NF-kB binding sites reduced the luciferase expression. Values are mean S.E.M. of three experiments run in triplicate p < 0.05 vs. the corresponding control value p < 0.05 vs. corresponding wild-type luciferase reporter plasmid). For methods and details, see Samico et al. (2009).

PLL-based carriers have been used as vehicles for DNA vector-based RNAi in combination with a multifunctional envelope-type nanodevice. This combination complexed to a DNA plasmid encoding anti-luciferase siRNA demonstrated 96% inhibition of luciferase activity in an in vitro co-transfection study (57). 

Fig. 2. TransIT-LTl-mediated transfection of HeLa cells at varying cell densities HeLa cells were plated in 12-well plates and transfected in parallel at 50% and 90% confluency. TransIT-LTl reagent transfections were performed in duplicate using a luciferase expression vector (pCI-luc) and 3 iL reagent per well, shows the importance of plating cells at optimal density for transfection. Twenty-four hours post-transfection, cells were harvested and assayed for luciferase activity. Visual confluence (line graph) was measured under the microscope at harvest. The data represent the average luciferase activity (Relative Light Units - RLUs in Millions) from three experiments performed on different days.

The authors would like to thank Dr. Bob Scholte for transduction of the H441cells with eGFP and luciferase using lentiviral vectors. This work was supported by the European Union through the Project FP6-LSHB-CT-2006-019038 Magselectofection, as well as by the German Ministry of Education and Research,... 

Cationic vesicles typically used for DNA delivery often self-aggregate or bind to plasma proteins in vivo. Wu et al. [104] attempted to improve vesicle stability using a cationic lipid with a cross-linkable acrylamide attached to the headgroup (Fig. 16). Vesicles were polymerized using thermal initiation with AAPD. Compared to monomeric vesicles, polymerized vesicles were less cytotoxic, more resistant to aggregation in serum, and comparable in transfection activity using a vector encoding firefly luciferase. 

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