pGEM4Z/A64 plasmid vector穩(wěn)定型體外轉(zhuǎn)錄mRNA疫苗載體質(zhì)粒

In recent years, improvements in plasmid vectors have increased the stability of the in vitro transcribed mRNA, turning our attention to mRNA drugs, especially to mRNA vaccines. pGEM4Z/GFP/A64 and pGEM4Z/OVA/A64 are constructed based on pGEM4Z/A64 vector and made as templates for producing the in vitro transcribed mRNAs, which are inoculated via the intranasal route to induce anti-tumor immunity [Phua K K, et al. Sci Rep. 2014; 4:5128]. Using pcDNA3.1-64A and pSP73-Sph/A64, several vectors containing tumor-associated antigens (TAA), glucocorticord-induced TNFR-related protein monoclonal antibody (GITR mAb) and cytotoxic T-lymphocyte-associated protein-4 mAb (CTLA-4 mAb) are respectively constructed and used for producing the corresponding in vitro transcribed mRNAs, which are electroporated into dendritic cells (DC). Subsequently the obtained DC-mRNA vaccines are used for enhancing anti-tumor immunity [Pruitt S K, et al. Eur J Immunol. 2011; 41(12): 3553-63]. pSpjC-βglacZβgan and pT7TSβggfpβgan are respectively constructed, resulting in LacZ and green fluorescent protein (GFP) genes flanked by 5′-untranslated region (UTR) and 3′UTR from xenopus laevis β-globin respectively [Hoerr I, et al. Eur J Immunol. 2000; 30 (1): 1-7]. The plasmid vectors containing TAA such as mucin1 (MUC1), carcinoembryonic antigen (CEA), human epidermal growth factor receptor 2 (Her-2/neu), telomerase, survivin and melanoma-associated antigen 1 (MAGE-1) are respectively constructed utilizing pSP64-Poly (A)-EGFP-2 provided by V.F.I. Van Tendeloo and taken as templates for producing the in vitro transcribed mRNAs, which are used for anti-tumor immunity [Rittig S M, et al. Mol Ther. 2011; 19 (5): 990-9]. Also 5′top UTR is artificially synthesized and applied for increasing mRNA stability [Andreas Thess. US 20150050302 A1. Artificial nucleic acid molecules comprising a 5′top utr]. Several plasmids containing multiple mutant major histocompatibility complex (MHC) class II epitope sequences are respectively constructed using pST1-Sp-MITD-2hBgUTR-A120 and used for producing the in vitro transcribed mRNAs, which are inoculated into the body for generating personalized anti-cancer immunity

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