Johanning F W, Conry R M, LoBuglio A F, Wright M, Sumerel L A, Pike M J, Curiel D T
Gene Therapy Program, University of Alabama at Birmingham 35294, USA.
Nucleic Acids Res. 1995 May 11;23(9):1495-501. doi: 10.1093/nar/23.9.1495.
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.
已证明直接肌内递送裸质粒DNA可使编码的异源基因在靶肌细胞中表达。该方法已被用于基于递送编码抗原的质粒DNA引发免疫。为了应用于实现针对抗原性转化癌蛋白的抗肿瘤免疫，递送编码这些分子的质粒DNA会产生重大的潜在安全隐患。作为DNA多核苷酸载体的替代方法，我们探索了mRNA载体在体内诱导肌肉细胞中外源基因表达的效用。为此分析获得了合成的编码报告基因的mRNA转录物。辛德毕斯病毒载体也被用于获得具有自我复制能力的荧光素酶mRNA转录物。在这些研究中，可以表明与非复制性mRNA种类相比，复制性载体能够在肌细胞中指导显著更高水平的报告基因表达。此外，与非复制性mRNA相比，复制性种类能够实现显著延长的体内基因表达水平。这两个特性将使复制性mRNA载体可用于基于多核苷酸的免疫方案。
