Maintenance and Gene Electrotransfer Efficiency of Antibiotic Resistance Gene-Free Plasmids Encoding Mouse, Canine and Human Interleukin-12 Orthologues
| العنوان: | Maintenance and Gene Electrotransfer Efficiency of Antibiotic Resistance Gene-Free Plasmids Encoding Mouse, Canine and Human Interleukin-12 Orthologues |
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| المؤلفون: | Urska Kamensek, Andrej Rencelj, Tanja Jesenko, Tinkara Remic, Gregor Sersa, Maja Cemazar |
| المصدر: | Heliyon, str. 1-08879-10-08879, Vol. 8, no. 2, Feb. 2022 COBISS-ID: 21607432 Heliyon, vol. 8, no. 2, pp. 1-08879-10-08879, 2022. Heliyon, Vol 8, Iss 2, Pp e08879-(2022) |
| بيانات النشر: | Elsevier BV, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | History, Science (General), Polymers and Plastics, interleukin-12, Antibiotic resistance gene-free, electrotransfer, interleukin-12, immunotherapy, mammals, Plasmid, Industrial and Manufacturing Engineering, Interleukin 12, udc:602.6/.7, elektroprenos genov, interlevkin 12, imunoterapija, sesalci, Q1-390, Gene electrotransfer, interlevkin 12, sesalci, elektroprenos genov, mammals, Business and International Management, electrotransfer, H1-99, Multidisciplinary, Copy number, imunoterapija, Transfection efficiency, Social sciences (General), immunotherapy |
| الوصف: | Interleukin 12 (IL-12) is a cytokine used as a therapeutic molecule in cancer immunotherapy. Gene electrotransfer mediated delivery of IL-12 gene has reached clinical evaluation in the USA using a plasmid that in addition to IL- 12 gene also carry an antibiotic resistance gene needed for its production in bacteria. In Europe however, Eu- ropean Medicines Agency recommends against the use of antibiotics during the production of clinical grade plasmids. We have prepared several antibiotic resistance gene-free plasmids using an antibiotic-free selection strategy called operator-repressor titration, including plasmids encoding mouse, canine and human IL-12 orthologues. The aim of this study was to evaluate the maintenance of these plasmids in bacterial culture and test their transfection efficiency using gene electrotransfer. Plasmid maintenance was evaluated by determining plasmid yields and topologies after subculturing transformed bacteria. Transfection efficiency was evaluated by determining the plasmid copy number, expression and cytotoxicity after gene electrotransfer to mouse, canine and human melanoma cells. The results demonstrated that our IL-12 plasmids without an antibiotic resistance gene are stably maintained in bacteria and provide sufficient IL-12 expression after in vitro gene electrotransfer; therefore, they have the potential to proceed to further in vivo evaluation studies. Nasl. z nasl. zaslona. Opis vira z dne 7. 2. 2022. Bibliografija na koncu prispevka. Abstract. |
| وصف الملف: | text/url; application/pdf |
| تدمد: | 1556-5068 2405-8440 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2f0fdebe2551d4b3ce6d16290189d4b6Test https://doi.org/10.2139/ssrn.3977521Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....2f0fdebe2551d4b3ce6d16290189d4b6 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15565068 24058440 |
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