A SINDBIS VIRUS MESSENGER-RNA POLYNUCLEOTIDE VECTOR ACHIEVES PROLONGED AND HIGH-LEVEL HETEROLOGOUS GENE-EXPRESSION IN-VIVO
A SINDBIS VIRUS MESSENGER-RNA POLYNUCLEOTIDE VECTOR ACHIEVES PROLONGED AND HIGH-LEVEL HETEROLOGOUS GENE-EXPRESSION IN-VIVO
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DOI：
10.1093/nar/23.9.1495
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发表时间：
1995-05-11
影响因子：
14.9
通讯作者：
CURIEL, DT
中科院分区：
文献类型：
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作者：
JOHANNING, FW;CONRY, RM;CURIEL, DT
裸质粒DNA的直接肌肉内传递已被证明允许编码的异源基因在靶细胞中表达。该方法已被用于诱导基于抗原编码质粒DNA的免疫。在实现针对抗原性转化癌蛋白的抗肿瘤免疫的应用中，编码这些分子的质粒DNA的传递将产生重大的潜在安全隐患。作为DNA多核苷酸载体的替代，我们探索了mRNA载体在体内诱导外源基因在肌肉细胞中表达的效用。合成的报告基因编码的mRNA转录本被用于这一分析。还利用Sindbis病毒载体获得了具有自我复制能力的荧光素酶mRNA转录本。在这些研究中，可以证明复制型载体能够引导报告基因在心肌细胞中的表达水平显著高于非复制型mRNA物种。此外，与非复制型mRNA相比，复制型物种能够显著延长体内基因表达水平。这两个特点都将使复制型mRNA载体成为基于多核苷酸的免疫方案的有用工具。
The direct intramuscular delivery of-naked plasmid DNA has been demonstrated to allow expression of encoded heterologous genes in the target myocytes. The method has been employed to elicit immunization based upon delivery of antigen encoding plasmid DNA. For application in the context of achieving anti-tumor immunization against antigenic transforming oncoproteins, delivery of plasmid DNAs encoding these molecules would create significant potential safety hazards. As an alternative to DNA polynucleotide vectors, we explored the utility of mRNA vehicles for inducing foreign gene expression in muscle cells in vivo. Synthetic reporter-gene encoding mRNA transcripts were derived for this analysis. The sindbis virus vector was also used to derive luciferase mRNA transcripts which possessed self-replication capacity. In these studies, it could be shown that the replicative vector was capable of directing significantly elevated levels of reporter gene expression in myocytes compared to a non-replicative mRNA species. In addition, the replicative species was capable of achieving significantly prolonged levels of in vivo gene expression compared to non-replicative mRNA. Both of these characteristics will make replicative mRNA vectors of utility for polynucleotide-based immunization protocols.