pGEM4Z/OVA/A64  plasmid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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