An effective gene-knockdown using multiple shRNA-expressing adenovirus vectors
An effective gene-knockdown using multiple shRNA-expressing adenovirus vectors
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使用多个表达 shRNA 的腺病毒载体进行有效的基因敲除
DOI：
10.1016/j.jconrel.2011.04.009
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发表时间：
2011
影响因子：
10.8
通讯作者：
et al
中科院分区：
文献类型：
--
作者：
Motegi Y;et al
表达短发夹RNA（shRNA）的病毒载体对于高效和组织特异性的RNA干扰（RNAi）递送是有吸引力的。我们和其他人先前报道了重组腺病毒（Ad）载体介导的RNAi在分子生物学研究和基因治疗中具有巨大的应用潜力。在本研究中，我们开发了一种有效的Ad载体介导的RNAi系统，其中Ad载体携带四个shRNA表达盒（Ad-multi-shRNA载体），这是一种简单有效的增强每个Ad载体颗粒的RNAi反应的策略。数据表明，与含有单个shRNA表达盒的常规Ad载体相比，Ad-多-shRNA载体显示出增强的RNAi效应。携带针对RET指蛋白（已知使细胞对氧化应激和顺铂脱敏的癌基因）的四个相同shRNA序列的Ad-多-shRNA载体的应用导致顺铂的细胞毒性作用增强，证明了Ad-多-shRNA载体用于沉默靶基因的优势。此外，携带四种不同shRNA序列的Ad-multi-shRNA有效地同时沉默多个靶基因。这些数据表明，Ad-multi-shRNA载体不仅在基础研究中，而且在临床基因治疗中的潜在用途。
Viral vectors expressing short hairpin RNA (shRNA) are attractive for efficient and tissue-specific RNA interference (RNAi) delivery. We and others previously reported that recombinant adenovirus (Ad) vector-mediated RNAi has great potential for a variety of applications in molecular biology studies and gene therapy. In the present study, we have developed an efficient Ad vector-mediated RNAi system, in which an Ad vector carries four shRNA-expression cassettes (Ad-multi-shRNA vector), a simple and effective strategy for enhancing the RNAi response per Ad vector particle. The data demonstrated that the Ad-multi-shRNA vectors showed an enhanced RNAi effect compared to conventional Ad vectors containing a single shRNA-expression cassette. An application of the Ad-multi-shRNA vector carrying four same shRNA-sequences against the RET finger protein, an oncogene known to desensitize cells to oxidative stress and cisplatin, resulted in an enhanced cytotoxic effect of cisplatin, demonstrating the advantages of the Ad-multi-shRNA vector for silencing target genes. Furthermore, an Ad-multi-shRNA carrying four different shRNA-sequences efficiently silenced the multiple target genes simultaneously. These data suggest the potential usefulness of the Ad-multi-shRNA vector not only in basic research but also in clinical gene therapy.