一个有针对性,自传递和光控制的分子信标,用于在活细胞中检测mRNA
Liping Qiu1, Cuichen Wu, Mingxu You
1Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, Hunan University, Changsha 410082, China.
Journal of the American Chemical Society
|August 13, 2013
概括
研究人员开发了一种基于光控制的aptamer的分子信标 (MB) 用于活细胞mRNA成像. 这种有针对性的探测器使细胞内mRNA动态的精确时空监测成为可能,克服了以前的检测挑战.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 在活细胞中成像和检测特定的mRNA分子对化学和生物医学研究提出了重大挑战.
- 现有的方法对于细胞内mRNA动态的时空分析缺乏精度.
研究的目的:
- 开发一种新型的分子信标 (MB),用于有针对性,自传递和光控制的细胞内mRNA分析.
- 为了实现精确的时空成像和检测活细胞内部的mRNA分子.
主要方法:
- 开发了一种基于aptamer的分子信标 (MB),与载体探头 (CP) 集成,用于细胞特异性传递.
- 使用内部化阿巴胺 (AS1411) 向细胞膜上的核素.
- 在CP中纳入光标基组,用于光激活释放和MB的功能.
- 采用CP作为一个追踪器和内部参考器,用于比度mRNA检测.
主要成果:
- 通过aptamer-nucleolin相互作用成功地证明了MB的针对性传递到特定的细胞.
- 实现了光控制的MB释放,从而实现了mRNA监测的精确时空激活.
- 验证了MB对活细胞mRNA成像具有高时空分辨率的能力.
- 展示了CP在跟踪探头分布和实现比度mRNA定量方面的双重作用.
结论:
- 开发的基于光控制的aptamer的MB系统为细胞内mRNA分析提供了强大的工具.
- 这项技术克服了空间时空mRNA成像的局限性,推进了化学和生物医学研究.
- 提供对mRNA检测的精确控制,促进对活细胞中基因表达动态的更深入的理解.
相关概念视频
Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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...
The real-time quantification of the number of amplified products is...
DNA Microarrays
Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...


