Maintenance and gene electrotransfer efficiency of antibiotic resistance gene-free plasmids encoding mouse, canine and human interleukin-12 orthologues

Kamenšek, Urška; Rencelj, Andrej; Jesenko, Tanja; Remic, Tinkara; Serša, Gregor; Čemažar, Maja

doi:10.1016/j.heliyon.2022.e08879

Cited by 3 publications

(2 citation statements)

References 69 publications

(95 reference statements)

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“…The pORT plasmids have mostly been used to design DNA vaccines. Preclinical tests were performed with a pORT plasmid expressing either the LACK antigen for vaccination against canine leishmaniasis [ 75 ] or the interleukin-12 protein electrotransferred to mouse, canine and human melanoma cells to pave the way for a human clinical trial [ 76 ].…”

Section: Plasmids Devoid Of Antibiotic Resistance Marker and Containi...mentioning

confidence: 99%

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

Marie,

Scherman

2024

Genes

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Until very recently, the major use, for gene therapy, specifically of linear or circular DNA, such as plasmids, was as ancillary products for viral vectors’ production or as a genetic template for mRNA production. Thanks to targeted and more efficient physical or chemical delivery techniques and to the refinement of their structure, non-viral plasmid DNA are now under intensive consideration as pharmaceutical drugs. Plasmids traditionally carry an antibiotic resistance gene for providing the selection pressure necessary for maintenance in a bacterial host. Nearly a dozen different antibiotic-free gene vectors have now been developed and are currently assessed in preclinical assays and phase I/II clinical trials. Their reduced size leads to increased transfection efficiency and prolonged transgene expression. In addition, associating non-viral gene vectors and DNA transposons, which mediate transgene integration into the host genome, circumvents plasmid dilution in dividing eukaryotic cells which generate a loss of the therapeutic gene. Combining these novel molecular tools allowed a significantly higher yield of genetically engineered T and Natural Killer cells for adoptive immunotherapies due to a reduced cytotoxicity and increased transposition rate. This review describes the main progresses accomplished for safer, more efficient and cost-effective gene and cell therapies using non-viral approaches and antibiotic-free gene vectors.

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Section: Plasmids Devoid Of Antibiotic Resistance Marker and Containi...mentioning

confidence: 99%

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

Marie,

Scherman

2024

Genes

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“…In the field of immunotherapy by gene transfer, most studies report electrotransfer of plasmid DNA encoding interleukin 12 (IL-12) [ 4 , 8 , 9 ]. Due to its potent pro-inflammatory functions IL-12 is a suitable candidate for cancer immunotherapy [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Gene Immunotherapy of Colon Carcinoma with IL-2 and IL-12 Using Gene Electrotransfer

Komel,

Omerzel,

Kamensek

et al. 2023

IJMS

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Gene immunotherapy has become an important approach in the treatment of cancer. One example is the introduction of genes encoding immunostimulatory cytokines, such as interleukin 2 and interleukin 12, which stimulate immune cells in tumours. The aim of our study was to determine the effects of gene electrotransfer of plasmids encoding interleukin 2 and interleukin 12 individually and in combination in the CT26 murine colon carcinoma cell line in mice. In the in vitro experiment, the pulse protocol that resulted in the highest expression of IL-2 and IL-12 mRNA and proteins was used for the in vivo part. In vivo, tumour growth delay and also complete response were observed in the group treated with the plasmid combination. Compared to the control group, the highest levels of various immunostimulatory cytokines and increased immune infiltration were observed in the combination group. Long-term anti-tumour immunity was observed in the combination group after tumour re-challenge. In conclusion, our combination therapy efficiently eradicated CT26 colon carcinoma in mice and also generated strong anti-tumour immune memory.

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Electroporation in Clinical Applications—The Potential of Gene Electrotransfer and Electrochemotherapy

et al. 2022

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Electroporation (EP) allows for the transport of molecules into the cytoplasm with significant effectiveness by forming transient pores in the cell membrane using electric pulses. This can be used for cellular transport (RE—reversible electroporation) or ablation (IRE—irreversible electroporation). The first of described options fortifies medicine with novel possibilities: electrochemotherapy (ECT), which creates promising perspectives for cancer treatment, and gene electrotransfer (GET), a powerful method of DNA delivery as well as immunogen electrotransfer. The review constitutes a comprehensive explanation of the mechanism of EP in the case of GET, its present and prospective employment in medicine, including gene delivery, vaccinations, therapy, and transfection, are also presented.

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Maintenance and gene electrotransfer efficiency of antibiotic resistance gene-free plasmids encoding mouse, canine and human interleukin-12 orthologues

Cited by 3 publications

References 69 publications

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

Gene Immunotherapy of Colon Carcinoma with IL-2 and IL-12 Using Gene Electrotransfer

Electroporation in Clinical Applications—The Potential of Gene Electrotransfer and Electrochemotherapy

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