A Sendai Virus-Based Cytoplasmic RNA Vector as a Novel Platform for Long-Term Expression of MicroRNAs
A Sendai Virus-Based Cytoplasmic RNA Vector as a Novel Platform for Long-Term Expression of MicroRNAs
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DOI：
10.1016/j.omtm.2019.10.012
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发表时间：
2019-10
期刊：
影响因子：
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通讯作者：
M. Sano;Asako Nakasu;M. Ohtaka;M. Nakanishi
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文献类型：
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作者：
M. Sano;Asako Nakasu;M. Ohtaka;M. Nakanishi
细胞质RNA病毒衍生的载体已经成为用于microRNA（miRNA）递送的有吸引力的载体，因为它们不具有染色体插入的潜在风险。然而，它们相对短期的表达限制了它们在需要长期miRNA操作的生物学应用中的使用，例如体细胞重编程。在这里，我们表明，基于复制缺陷型和持续性仙台病毒（SeVdp）的细胞质RNA病毒载体可作为长期生产能够诱导序列特异性靶抑制的miRNA的有效平台。SeVdp载体能够同时将富含胚胎干细胞的miRNA以及多种转录因子递送到成纤维细胞中，从而有效地重编程为诱导多能干细胞。此外，我们报告说，小鼠miR-367发夹产生的成熟miRNA的水平升高时，它被纳入SeVdp载体，并作为一个有效的骨架生产人工miRNA。这些SeVdp载体衍生的人工miRNA有效地抑制靶基因的表达。我们的发现为再生医学、基因治疗和细胞治疗等领域的长期和靶向基因沉默提供了新的见解。
Cytoplasmic RNA virus-derived vectors have emerged as attractive vehicles for microRNA (miRNA) delivery as they possess no potential risk of chromosomal insertion. However, their relatively short-term expression limits their use in biological applications that require long-term miRNA manipulation, such as somatic cell reprogramming. Here, we show that a cytoplasmic RNA virus vector based on a replication-defective and persistent Sendai virus (SeVdp) serves as an effective platform for long-term production of miRNAs capable of inducing sequence-specific target suppression. The SeVdp vector was able to simultaneously deliver embryonic stem cell-enriched miRNAs, as well as multiple transcription factors, into fibroblasts, resulting in effective reprogramming into induced pluripotent stem cells. Furthermore, we report that the murine miR-367 hairpin produced elevated levels of mature miRNA when it was incorporated into the SeVdp vector and served as an effective backbone for production of artificial miRNAs. These SeVdp vector-derived artificial miRNAs efficiently inhibited expression of target genes. Our findings provide novel insights into a powerful tool for long-term and targeted gene silencing in areas such as regenerative medicine, gene therapy, and cell therapy.