Spliced spleen necrosis virus vector RNA is not encapsidated: implications for retroviral replication and vector design.
Spliced spleen necrosis virus vector RNA is not encapsidated: implications for retroviral replication and vector design.
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剪接的脾坏死病毒载体 RNA 未衣壳化：对逆转录病毒复制和载体设计的影响。
DOI：
10.1016/j.ymthe.2004.01.009
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Pomerantz,RogerJ
中科院分区：
文献类型：
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作者：
Goodrich,Adrienne;Parveen,Zahida;Dornburg,Ralph;Schnell,MatthiasJ;Pomerantz,RogerJ
RNA剪接是逆转录病毒生命周期中的一个复杂事件，可能涉及多个步骤以及升序作用序列，以维持剪接和非剪接病毒RNA的翻译和封装的适当平衡。逆转录病毒RNA可以由细胞机器处理，并能够去除内含子序列。我们的目标是利用去除合成内含子来进行靶向基因表达。为了分析内含子去除和基因表达，我们构建了一种新型的自失活基因激活(SIGA)载体，用于潜在的普遍基因治疗。新的基因治疗载体对于人类安全有效的基因传递是必要的。SIGA载体来源于脾坏死病毒(SNV)，SNV是一种禽网状内皮组织增生症病毒。该载体的设计使得治疗基因的表达在辅助细胞系中被阻止，因为中间序列包含转录和翻译中的各种区块。然而，在一轮逆转录病毒复制之后，介入序列应该被细胞机器移除，治疗基因将在靶细胞中选择性地表达。我们的研究表明，SIGA载体RNA中的插入序列被部分剪接。然而，剪接载体RNA没有被转导到靶细胞。以前的研究表明，具有感染性的SNV载体能够转导剪接的RNA。然而，感染性和复制缺陷逆转录病毒复制的未知差异可能会对剪接RNA的转导产生影响。这项研究的结果提供了有关剪接RNA及其包裹的关键信息，以及构建新一代SNV衍生逆转录病毒载体的数据。
RNA splicing is a complex event in the retroviral life cycle and can involve multiple steps, as well ascis-acting sequences, to maintain a proper balance of spliced and unspliced viral RNA for translation and encapsidation. The retroviral RNA can be processed by cellular machinery and enables the removal of intronic sequences. We aimed to utilize the removal of a synthetic intron for targeted gene expression. To analyze intron removal and gene expression, we have constructed a novel self-inactivating gene-activating (SIGA) vector for potential universal gene therapy. New vectors for gene therapy are necessary for safe and effective gene delivery in humans. The SIGA vector is derived from spleen necrosis virus (SNV), which is an avian reticuloendotheliosis virus. The vector was designed so that expression of a therapeutic gene is blocked in helper cell lines due to an intervening sequence containing various blocks in transcription and translation. However, after one round of retroviral replication, the intervening sequence should be removed by the cellular machinery and the therapeutic gene will be selectively expressed in target cells. Our studies show that the intervening sequence in SIGA vector RNA is partially spliced. However, spliced vector RNA was not transduced to target cells. Previous studies showed that an infectious SNV vector enabled transduction of spliced RNA. However, yet-undefined differences in infectious and replication-deficient retroviral replication may have an effect on the transduction of spliced RNA. The results of this study present key information on spliced RNA and its encapsidation, as well as data for the construction of a new generation of SNV-derived retroviral vectors.