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.

中文翻译：

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基于仙台病毒的细胞质RNA载体作为MicroRNA长期表达的新型平台。

细胞质RNA病毒衍生的载体已成为microRNA（miRNA）传递的诱人载体，因为它们没有染色体插入的潜在风险。但是，它们相对短期的表达限制了它们在需要长期操作miRNA（例如体细胞重编程）的生物学应用中的使用。在这里，我们显示基于复制缺陷和持久性仙台病毒（SeVdp）的细胞质RNA病毒载体可作为长期生产miRNA的有效平台，能够诱导序列特异性靶点抑制。SeVdp载体能够将富含胚胎干细胞的miRNA以及多种转录因子同时递送到成纤维细胞中，从而有效地重编程为诱导性多能干细胞。此外，我们报告说，将鼠miR-367发夹整合到SeVdp载体中后，其产生的成熟miRNA的水平升高，并作为生产人造miRNA的有效骨架。这些来自SeVdp载体的人工miRNA可有效抑制靶基因的表达。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的强大工具提供了新颖的见解。