Tailor-made fibroblast-specific and antibiotic-free interleukin 12 plasmid for gene electrotransfer-mediated cancer immunotherapy

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• 作者：Urska Kamensek^a ; ^{ukamensek@onko-i.si} ; Natasa Tesic^b ; ^{natasa.tesic@fvz.upr.si} ; Gregor Sersa^a ; ^{gsersa@onko-i.si} ; Spela Kos^a ; ^{skos@onko-i.si} ; Maja Cemazar^a ; ^b ; ^{mcemazar@onko-i.si}
• 关键词：IL-12 ; interleukin 12 ; GET ; gene electrotransfer ; CMV ; cytomegalovirus ; EMA ; European medicines agency ; ORT ; operator-repressor titration ; SVEC4&ndash ; 10 ; murine endothelial cells ; HUVEC ; human umbilical vein endothelial cells ; L929 ; murine fibroblasts ; LPB ; murine fibrosarcoma cells ; WI-38 ; human fibroblasts ; GFP ; green fluorescent protein ; EF-1α/HTLV ; hybrid elongation factor-1α promoter ; ELISA ; enzyme-linked immunosorbent assay ; qRT-PCR ; quantitative reverse transcription polymerase chain rea
• 刊名：Plasmid
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：89
• 期：Complete
• 页码：9-15
• 全文大小：1063 K
• 卷排序：89

文摘

An optimized IL-12 plasmid intended for GET mediated cancer gene therapy was constructed. The ubiquitous promoter was replaced with a promoter of the collagen gene for fibroblast-specific expression. The plasmid was prepared using an antibiotic-free technology to comply with the regulatory demands for clinical use. The constructed fibroblast-specific and antibiotic-free IL-12 plasmid supports low expression after GET. The removal of antibiotic resistance did not affect the expression profile of the plasmid and lowered its cytotoxicity.