相关实验视频

Updated: May 21, 2025

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Published on: September 13, 2013

模块化访问核基 GFP 替代品:适应 pH 的智能探针用于比值核酸诊断

Keenan T Regan¹, Austin Pounder², Ryan E Johnson¹

¹Department of Chemistry & Toxicology, University of Guelph Guelph Ontario N1G 2W1 Canada rmanderv@uoguelph.ca.

Chemical science

|March 19, 2025

概括

研究人员开发了模仿GFP染色体的新型光分子转子 (FMR),用于核酸诊断. 这些FMR显示了增强的亮度和开启响应,使得DNA和G-四重复结构中的敏感检测和pH监测成为可能.

科学领域:

超分子化学超分子化学
生物物理化学生物物理化学
核酸化学的核酸化学

背景情况:

目前用于核酸应用的绿色光蛋白 (GFP) 替代品存在低亮度和弱开启反应.
模仿氨酸 (Tyr66) 染色体4-基基基基胺 (HBI) 对于开发高效的GFP替代品至关重要.
光分子转子 (FMR) 通过扭曲的分子内电荷转移 (TICT) 提供了增强光物理性能的潜力.

研究的目的:

合成基于石墨板的FMR作为具有更好的亮度和响应能力的新型GFP替代品.
将这些FMR纳入含有寡核酸的DNA和锁定核酸 (LNA) 中 (双重复和G-四重复).
在各种环境中研究这些FMR修饰核酸的结构和光物理特性.

主要方法:

基于基的FMRs的模块化链上阿尔多尔合成.
将FMR纳入具有正规和LNA基的NarI12和TBA15寡核酸中.
使用光谱学,分子动力学模拟和量子力学计算进行表征.

主要成果:

FMR显示了可调节的光物理性质,富含电子的石墨板显示双排放和缺乏电子的石墨板作为比率指标.
在使用LNA的G-四倍到双倍拓切换器中观察到显著的启动响应 (高达154倍).
优化的FMR在双重体中实现了高光量子产量 (Φ_fl高达0.58) 和光亮度 (∼15,000 cm^-1 M^-1) .
一个可切换的光诱导电子转移机制使得对pH值变化产生了134倍的开启发射反应.

结论:

开发的基于基的FMRs代表了首次成功优化GFP替代品作为内部核基替代品.
这些FMR在核酸环境中表现出优越的亮度和响应能力,与之前的GFP替代品相比.
这些发现表明,这些FMR是核酸诊断的有希望的工具,提供敏感的检测和pH监测能力.

更多相关视频

Synthesis of Wavelength-shifting DNA Hybridization Probes by Using Photostable Cyanine Dyes

Synthesis of Wavelength-shifting DNA Hybridization Probes by Using Photostable Cyanine Dyes

Published on: July 6, 2016

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Published on: April 9, 2018

相关概念视频

Labeling DNA Probes

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...

Reporter Genes

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.