头DNA功能化黄金合物用于活细胞中mRNA的成像
Ashwath Jayagopal1, Kristin C Halfpenny, Jonas W Perez
1Department of Chemistry, Vanderbilt University, Nashville, Tennessee 37235, USA.
Journal of the American Chemical Society
|July 1, 2010
概括
研究人员开发了头DNA功能化黄金纳米粒子 (hAuNP) 用于活细胞对信使RNA (mRNA) 的成像. 这种新的纳米粒子方法为研究RNA贩运和细胞过程提供了敏感和特定的mRNA检测.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 纳米技术 纳米技术
背景情况:
- 研究活细胞中的信使RNA (mRNA) 贩运对于理解基因表达和疾病至关重要.
- 现有的mRNA成像方法通常需要转染试剂,具有有限的特异性,或表现出高细胞毒性.
研究的目的:
- 开发一种基于纳米粒子的新策略,用于对mRNA敏感和特定的活细胞成像.
- 为了实现RNA贩运和下游分析的实时空间和时间研究.
主要方法:
- 使用头DNA功能化黄金纳米粒子 (hAuNP),包含用于目标mRNA识别的分子信标.
- 在没有转染试剂的情况下,证明了hAuNP的高效细胞内化.
- 评估了DNase耐药性,低细胞毒性和对目标mRNA序列 (GADPH和RSV mRNA) 的高特异性.
- 在活细胞中对不同的mRNA进行多重光成像,并在固定细胞中同时进行mRNA-蛋白质成像.
主要成果:
- hAuNP对目标mRNA具有高的特异性和敏感性,具有高的目标与背景比率.
- 在HEp-2细胞中成功对GADPH mRNA和感染HEp-2细胞中的RSV mRNA进行活细胞成像.
- 展示了多重和同时成像的能力.
- 药理学分析表明细胞内部化通过膜-纳米粒子相互作用.
结论:
- hAuNP代表了对mRNA活细胞成像的一个有前途的工具,克服了当前技术的局限性.
- 这种方法促进了mRNA运输和处理的实时分析,有助于阐明生物过程和疾病病原性.
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...
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.
Commonly used reporter...
Commonly used reporter...
Immunogold Electron Microscopy
Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.


