对siRNA非种子区域的干扰微调了沉默能力
Hanna K Hedman1, Finn Kirpekar, Sofi K C Elmroth
1Department of Biochemistry and Structural Biology, Center for Molecular Protein Science, Lund University, P.O. Box 124, SE-22100 Lund, Sweden.
Journal of the American Chemical Society
|July 5, 2011
概括
抗癌药物西斯和氧沙被用于化小干扰RNA (siRNAs). 化改变了siRNA降解,但保留了生物活性,尽管它通常降低了沉默能力,而氧化具有更大的效果.
科学领域:
- 分子生物学分子生物学
- RNA干扰 (RNAi) 是一种RNA干扰.
- 药物开发 药物开发
背景情况:
- 非编码RNAs,包括小干扰RNAs (siRNAs) 和microRNAs (miRNAs),在基因表达调节中起着至关重要的作用.
- siRNAs用于基因沉默,系统稳定性是临床应用的关键因素.
- 了解化疗药物等因素如何影响siRNA和miRNA功能至关重要,特别是在病理条件下.
研究的目的:
- 调查西斯和氧沙对siRNAs基因沉默功能的影响.
- 描述siRNAs上形成的-RNA附加物及其对RNA稳定性和功能的影响.
- 为了比较西斯与氧沙对siRNA处理和沉默能力的影响.
主要方法:
- 针对Wnt-5a mRNA (NM_003352) 的siRNAs的构建.
- 用西斯或氧沙先化反感性siRNA链.
- 使用凝电泳和MALDI-MS.检测和表征 adducts.
- 使用 luciferase 记者系统评估 HB2 细胞的沉默能力.
主要成果:
- 化siRNAs导致了改变降解模式的添加物,为修改点附近的水解提供了保护.
- MALDI-MS成功地识别和表征了化RNA,利用的天然同位素模式作为指纹.
- 反意义化siRNAs保留了生物活性,其化部位位于关键种子区域之外.
- 对于化siRNAs,观察到降低沉声能力的一般趋势.
- 与西斯相比,牛利对siRNA沉默能力产生了更大的影响.
结论:
- 抗癌药物可以修改siRNAs,影响它们的稳定性和基因沉默能力.
- 该研究提供了对基化疗药物与RNAi机制潜在干扰的见解.
- 西斯和氧沙在干扰siRNA和miRNA处理的能力方面存在微妙的差异.
相关概念视频
RNA Interference
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
RNA Interference
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
siRNA - Small Interfering RNAs
Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
Experimental RNAi
RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
Small interfering RNAs (siRNA)
Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
MicroRNAs
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...

