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纳秒脉冲激光诱导激波高效传递小干扰 RNA 至植物细胞用于转录后基因沉默。

Efficient delivery of small interfering RNA to plant cells by a nanosecond pulsed laser-induced stress wave for posttranscriptional gene silencing.

机构信息

Department of Biology, Howell Science Complex, East Carolina University, Greenville, NC 27858-4353, USA.

出版信息

Plant Sci. 2006 Sep;171(3):375-81. doi: 10.1016/j.plantsci.2006.04.005. Epub 2006 May 15.

Abstract

Small interfering RNA (siRNA) induced posttranscriptional gene silencing (PTGS) has been an efficient method for genetic and molecular analysis of certain developmental and physiological processes and represented a potential strategy for both controlling virus replication and developing therapeutic products. However, there are limitations for the methods currently used to deliver siRNA into cells. We report here, to our knowledge, the first efficient delivery of siRNA to plant cells by a nanosecond pulsed laser-induced stress wave (LISW) for posttranscriptional gene silencing. Using LISW, we are able to silence gene expression in cell cultures of three different plant species rice (Oryza sativa L.), cotton (Gossypium hirsutum L.), and slash pine (Pinus elliottii Engelm.). Gene silencing induced by siRNA has been confirmed by northern blot, laser scanning microscopy, and siRNA analysis. These data suggested that LISW-mediated siRNA delivery can be a reliable and effective method for inducing PTGS in cultured cells.

摘要

小干扰 RNA(siRNA)诱导的转录后基因沉默(PTGS)已成为研究某些发育和生理过程的遗传和分子的有效方法,并且代表了控制病毒复制和开发治疗产品的潜在策略。然而,目前用于将 siRNA 递送至细胞的方法存在局限性。我们在此报告,据我们所知,首次通过纳秒级脉冲激光诱导激波(LISW)有效地将 siRNA 递送至植物细胞,以实现转录后基因沉默。使用 LISW,我们能够沉默三种不同植物物种(水稻(Oryza sativa L.)、棉花(Gossypium hirsutum L.)和湿地松(Pinus elliottii Engelm.))的细胞培养物中的基因表达。通过 northern blot、激光扫描显微镜和 siRNA 分析证实了 siRNA 诱导的基因沉默。这些数据表明,LISW 介导的 siRNA 递送至培养细胞中的 PTGS 是一种可靠且有效的方法。

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