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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Retroviral delivery of small interfering RNA into primary cells
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Retroviral delivery of small interfering RNA into primary cells

机译:小干扰RNA逆转录病毒递送至原代细胞

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摘要

RNA interference is an evolutionarily conserved process in which recognition of double-stranded RNA ultimately leads to posttran-scriptional suppression of gene expression. This suppression is mediated by short (21- to 22-nt) small interfering RNAs (siRNAs), which induce degradation of mRNA based on complementary base pairing. The silencing of gene expression by siRNAs is emerging rapidly as a powerful method for genetic analysis. Recently, several groups have reported systems designed to express siRNAs in mammalian cells through transfection of either oligonucleotides or plasmids encoding siRNAs. Because these systems rely on transfection for delivery, the cell types available for study are restricted generally to transformed cell lines. Here, we describe a retroviral system for delivery of siRNA into cells. The use of retroviral vectors can greatly expand the types of cells available for RNA interference analysis. Furthermore, we demonstrate that this retroviral system allows for stable inactivation of genes in primary cells.
机译:RNA干扰是一个进化保守的过程,其中双链RNA的识别最终导致转录后的基因表达抑制。这种抑制作用是由短的(21至22 nt)小干扰RNA(siRNA)介导的,它们会基于互补碱基配对诱导mRNA降解。 siRNA沉默基因表达正在迅速成为一种强大的遗传分析方法。最近,一些研究小组报告了设计用于通过转染寡核苷酸或编码siRNA的质粒在哺乳动物细胞中表达siRNA的系统。因为这些系统依赖转染进行递送,所以可用于研究的细胞类型通常仅限于转化的细胞系。在这里,我们描述了将siRNA传递到细胞中的逆转录病毒系统。逆转录病毒载体的使用可以大大扩展可用于RNA干扰分析的细胞类型。此外,我们证明了这种逆转录病毒系统可以使原代细胞中的基因稳定失活。

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