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A Sendai Virus-Based Cytoplasmic RNA Vector as a Novel Platform for Long-Term Expression of MicroRNAs
Masayuki Sano1, Asako Nakasu1, Manami Ohtaka1
1Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 5, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
Abstract:
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.
Insights
A novel Sendai virus vector (SeVdp) enables long-term microRNA (miRNA) production for gene silencing. This platform effectively reprograms cells and shows promise for regenerative medicine and gene therapy applications.
Area of Science:
- Biotechnology
- Molecular Biology
- Virology
Background:
- Cytoplasmic RNA virus vectors offer safe microRNA (miRNA) delivery without chromosomal insertion risks.
- Short-term expression of current vectors limits applications requiring sustained miRNA activity, like cell reprogramming.
Purpose of the Study:
- To develop a replication-defective Sendai virus vector (SeVdp) for persistent miRNA production.
- To evaluate SeVdp's efficacy in long-term gene silencing and somatic cell reprogramming.
Main Methods:
- Engineered a replication-defective Sendai virus (SeVdp) vector for miRNA expression.
- Delivered miRNAs and transcription factors into fibroblasts using the SeVdp vector.
- Assessed miRNA production, gene silencing, and induced pluripotent stem cell (iPSC) generation.
Main Results:
- SeVdp enabled long-term miRNA production and sequence-specific target suppression.
- Simultaneous delivery of miRNAs and transcription factors via SeVdp effectively reprogrammed fibroblasts into iPSCs.
- Murine miR-367 hairpin backbone facilitated elevated artificial miRNA production within the SeVdp vector.
Conclusions:
- SeVdp is a powerful platform for sustained miRNA delivery and gene silencing.
- This technology holds significant potential for applications in regenerative medicine, gene therapy, and cell therapy.
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