Use and comparison of different internal ribosomal entry sites (IRES) in tricistronic retroviral vectors.
Use and comparison of different internal ribosomal entry sites (IRES) in tricistronic retroviral vectors.
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三顺反子逆转录病毒载体中不同内部核糖体进入位点（IRES）的使用和比较。
DOI：
10.1186/1472-6750-4-16
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发表时间：
2004-07-27
影响因子：
3.5
通讯作者：
Couderc, B
中科院分区：
文献类型：
--
作者：
Douin, V;Bornes, S;Creancier, L;Rochaix, P;Favre, G;Prats, AC;Couderc, B
多顺反子逆转录病毒载体含有通过内部核糖体进入位点（IRES）连接的多个治疗基因，为临床基因治疗方案中的外源cDNA的共表达提供了新的和有效的工具。例如，三顺反子逆转录病毒载体可用于遗传修饰抗原呈递细胞，使它们能够表达已知增强肿瘤细胞免疫原性的不同共刺激分子。我们已经构建并比较了不同的逆转录病毒载体，其含有两种共刺激分子（CD 70、CD 80）和连接到不同IRES序列（来自EMCV、c-myc、FGF-2和HTLV-1的IRES）的选择标记基因。含有来自EMCV、FGF-2或HTLV-1的IRES的三顺反子重组嗜异性病毒在诱导转导的鼠或人细胞中外源基因的表达方面同样有效，而不显示任何细胞类型特异性。然而，在同一mRNA上同时存在几个IRES可以诱导各种顺反子的差异表达。在这里，我们表明，在小鼠黑色素瘤细胞中，HTLV-1和EMCV的IRES干扰其他IRES诱导的翻译。然而，来自FGF-2的IRES确实诱导人黑素瘤细胞中外源cDNA的表达，而没有来自载体内存在的其它IRES的任何正或负调节。用三顺反子逆转录病毒载体遗传修饰的肿瘤细胞能够在鼠模型中诱导体内抗肿瘤免疫应答。外源基因的翻译由IRES指导，其高水平表达不仅取决于转导的细胞类型，还取决于载体内其他遗传元件的存在。
Polycistronic retroviral vectors that contain several therapeutic genes linked via internal ribosome entry sites (IRES), provide new and effective tools for the co-expression of exogenous cDNAs in clinical gene therapy protocols. For example, tricistronic retroviral vectors could be used to genetically modify antigen presenting cells, enabling them to express different co-stimulatory molecules known to enhance tumor cell immunogenicity. We have constructed and compared different retroviral vectors containing two co-stimulatory molecules (CD70, CD80) and selectable marker genes linked to different IRES sequences (IRES from EMCV, c-myc, FGF-2 and HTLV-1). The tricistronic recombinant amphotropic viruses containing the IRES from EMCV, FGF-2 or HTLV-1 were equally efficient in inducing the expression of an exogenous gene in the transduced murine or human cells, without displaying any cell type specificity. The simultaneous presence of several IRESes on the same mRNA, however, can induce the differential expression of the various cistrons. Here we show that the IRESes of HTLV-1 and EMCV interfere with the translation induced by other IRESes in mouse melanoma cells. The IRES from FGF-2 did however induce the expression of exogenous cDNA in human melanoma cells without any positive or negative regulation from the other IRESs present within the vectors. Tumor cells that were genetically modified with the tricistronic retroviral vectors, were able to induce an in vivo anti-tumor immune response in murine models. Translation of the exogenous gene is directed by the IRES and its high level of expression not only depends on the type of cell that is transduced but also on the presence of other genetic elements within the vector.
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影响因子：
4.4
作者：
Douin-Echinard, V;Péron, JM;Couderc, B
通讯作者：
Couderc, B
DOI：
10.1073/pnas.90.16.7642
发表时间：
1993-08-15
影响因子：
11.1
作者：
HELLEN, CUT;WITHERELL, GW;WIMMER, E
通讯作者：
WIMMER, E
影响因子：
11.4
作者：
BELSHAM, GJ
通讯作者：
BELSHAM, GJ
影响因子：
14.9
作者：
Bonnal, S;Boutonnet, C;Vagner, S
通讯作者：
Vagner, S
影响因子：
14.9
作者：
Borman, AM;LeMercier, P;Kean, KM
通讯作者：
Kean, KM