In vivo electroporation and stable transformation of skin cells of newborn mice by plasmid DNA

Titomirov, Alexander V.; Sukharev, Sergei; Kistanova, Elena

doi:10.1016/0167-4781(91)90162-f

Cited by 378 publications

(187 citation statements)

References 20 publications

(14 reference statements)

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“…1,2 It is now recognized as one of the most promising alternatives to viral vectors for transfection of different tissues in vivo for therapeutic purposes. [3][4][5] Gene therapy using electropulsation as a gene delivery method has already entered clinical trials for the treatment of different tumor types in cancer patients and for vaccination.…”

Section: Introductionmentioning

confidence: 99%

Control by pulse parameters of DNA electrotransfer into solid tumors in mice

et al. 2009

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Electrotransfer (electroporation) is recognized as one of the most promising alternatives to viral vectors for transfection of different tissues in vivo for therapeutic purposes. We evaluated the transfection efficiency of reporter genes (green fluorescent protein and luciferase) in murine subcutaneous tumors using different combinations of high-field (HV) (600-1400 V cm À1 , 100 ms, 8 pulses) and low-field (LV) (80-160 V cm À1 , 50-400 ms, 1-8 pulses) pulses and compared it to protocol using eight identical pulses of 600 V cm À1 and 5 ms duration (electro-gene therapy, EGT). Expression of GFP was determined using a fluorescent microscope and flow cytometry and expression of luciferase by measuring its activity using a luminometer. The EGT protocol yielded the highest expression of both reporter genes. However, a careful optimization of combinations of HV and LV pulses may result in similar transfection as EGT pulses. With the combination protocol, relatively high fields of LV pulses were necessary to obtain comparable transfection to the EGT protocol. Expression of reporter genes was higher in B16 melanoma than in SA-1 fibrosarcoma. Our data support the hypothesis that both electropermeabilization and electrophoresis are involved in electrotransfer of plasmid DNA, but demonstrate that these components have to happen at the same time to obtain significant expression of the target gene in tumors.

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Section: Introductionmentioning

confidence: 99%

Control by pulse parameters of DNA electrotransfer into solid tumors in mice

et al. 2009

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“…In the last 10 years, electric fields, with high intensity and short duration, have been used in vivo to permeabilize the plasma membrane of cells (ie electroporation) for delivery of plasmid DNA and cytotoxic agents. [2][3][4][5] Clinical trials of electric field-mediated drug delivery have been conducted to treat tumors. 6,7 These studies demonstrate that it is feasible and safe to use electric fields to facilitate drug and gene delivery.…”

Section: Introductionmentioning

confidence: 99%

Electromobility of plasmid DNA in tumor tissues during electric field-mediated gene delivery

Zaharoff

et al. 2002

Gene Ther

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“…This is due to the fact that many human or rodent tumors cannot be infected efficiently with adenoviral vectors due to lack of proper receptors. On the other hand, gene transfer by in vivo electroporation, a procedure that involves DNA injection followed by application of electric fields, is effective for introducing DNA into mouse muscle, 26 skin, 27 rat liver, 28 and tumors. 19,29 Earlier data show that use of in vivo electroporation enhances plasmid DNA uptake in tumor tissue, resulting in expression within the tumor.…”

Section: Discussionmentioning

confidence: 99%

Electroporation-mediated and EBV LMP1-regulated gene therapy in a syngenic mouse tumor model

Hsieh

Wu²,

Chow

et al. 2003

Cancer Gene Ther

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Latent membrane protein 1 (LMP1) is an Epstein-Barr virus (EBV)-encoded oncogene expressed in EBV-associated nasopharyngeal carcinoma (NPC). Previous studies indicate that a strategy combining LMP1-mediated NF-kB activation and the HSV thymidine kinase/Ganciclovir (HSVtk/GCV) prodrug system leads to regression of tumor growth in nude mice. To improve the efficacy of this strategy in immunocompetent hosts, we developed a therapeutic cassette, p6kB-EDL1E-tk, containing six copies of the NF-kB binding motif and an epithelial-specific EBV promoter, ED-L1E. The cassette was tested in a murine CT-26 carcinoma model in syngenic Balb/c mice. Coinjection of an LMP1-expressing vector and p6kB-EDL1E-tk by in vivo electroporation in mouse muscle revealed at least two-fold higher TK enzymatic activity than that of previously tested pLTR-tk. Furthermore, growth was attenuated in a group of mice containing LMP1-positive tumors that were intratumorally injected with the p6kB-EDL1E-tk cassette and GCV via in vivo electroporation, but not in mice treated with p6kB-EDL1E-tk or GCV alone. Similarly, no retardation of tumor growth was observed in mice containing LMP1-negative CT-26 tumors injected with both the p6kB-EDL1E-tk cassette and GCV. We propose that intratumoral injection of therapeutic agents, such as DNA of transcription-regulated cassette and GCV, via in vivo electroporation may be used as an alternative treatment for EBV LMP1-expressing cancers.

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In vivo electroporation and stable transformation of skin cells of newborn mice by plasmid DNA

Cited by 378 publications

References 20 publications

Control by pulse parameters of DNA electrotransfer into solid tumors in mice

Control by pulse parameters of DNA electrotransfer into solid tumors in mice

Electromobility of plasmid DNA in tumor tissues during electric field-mediated gene delivery

Electroporation-mediated and EBV LMP1-regulated gene therapy in a syngenic mouse tumor model

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