基于全方位的aptamer实时监测酶性RNA合成
Katharina Höfer1, Lisa V Langejürgen, Andres Jäschke
1Institute of Pharmacy and Molecular Biotechnology, Heidelberg University , 69120 Heidelberg, Germany.
Journal of the American Chemical Society
|September 3, 2013
概括
这项研究引入了一种通用光模块 (UFM),用于实时监测体外转录期间的RNA合成. 这种新的系统能够准确量化和评估任何合成的RNA的质量.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 合成生物学 合成生物学
背景情况:
- 在体外转录对于酶性RNA合成至关重要.
- 目前的方法缺乏在转录期间对RNA质量和数量的实时分析.
- 堕胎转录副产品使准确的监测变得复杂.
研究的目的:
- 开发一种通用系统,用于实时监测和定量任何RNA合成.
- 通过避免副产品的干扰来克服现有方法的局限性.
- 为了实现RNA合成的标准化和高通量选.
主要方法:
- 设计了一种包含菜阿普坦和子头 рибо酶的通用光模块 (UFM).
- 该UFM被添加到感兴趣的RNA (ROI).
- 主要转录的切割释放了UFM,它结合DFHBI染料产生光.
主要成果:
- 该系统提供RNA合成的实时量化,独立于RNA序列,聚合酶或促进体.
- 该UFM允许监测没有干扰从堕胎的转录产品.
- 证明了标准化,并行化和高通量选的潜力.
结论:
- 开发的通用光模块为实验室RNA合成的实时分析提供了强大而通用的解决方案.
- 这种方法显著提高了评估RNA生产质量和数量的能力.
- 促进RNA合成应用中的高效优化和选.
相关概念视频
Real Time RT-PCR
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...


