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Use of adeno-associated viral vector for delivery of small interfering RNA

机译：腺相关病毒载体在传递小分子干扰RNA中的应用

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摘要

Post-transcriptional gene silencing by small interfering RNAs (siRNAs) is rapidly becoming a powerful tool for genetic analysis of mammalian cells. Delivery of siRNA into mammalian cells is usually achieved via the transfection of double-stranded oligonucleotides or plasmids encoding RNA polymerase III promoter-driven small hairpin RNA. Recently, retroviral vectors have been used for siRNA delivery, which overcome the problem of poor transfection efficiency seen with the plasmid-based systems. However, retroviral vectors have several limitations, such as the need for active cell division for gene transduction, oncogenic potential, low titers and gene silencing. In this report, we have adapted a commercially available adenoassociated virus (AAV) vector for siRNA delivery into mammalian cells. We demonstrate the ability of this modified vector to deliver efficiently siRNA into HeLa S3 cells and downregulate p53 and caspase 8 expression. Our results suggest that AAV-based vectors are efficient vectors for the delivery of siRNA into mammalian cells. Based on the known ability of these vectors to infect both dividing and nondividing cells, their use as a therapeutic tool for the delivery of siRNA deserves further study.
机译：小干扰RNA（siRNA）的转录后基因沉默正在迅速成为哺乳动物细胞遗传分析的有力工具。 siRNA传递到哺乳动物细胞中通常是通过转染双链寡核苷酸或编码RNA聚合酶III启动子驱动的小发夹RNA的质粒实现的。最近，逆转录病毒载体已用于siRNA递送，克服了基于质粒的系统所见的转染效率差的问题。然而，逆转录病毒载体具有若干局限性，例如需要基因转导的活性细胞分裂，致癌潜力，低滴度和基因沉默。在本报告中，我们将可商购的腺伴随病毒（AAV）载体用于siRNA传递到哺乳动物细胞中。我们证明了这种修饰的载体能够有效地将siRNA传递到HeLa S3细胞中并下调p53和caspase 8的表达。我们的结果表明，基于AAV的载体是将siRNA传递到哺乳动物细胞中的有效载体。基于这些载体感染分裂细胞和非分裂细胞的已知能力，将其用作递送siRNA的治疗工具值得进一步研究。

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