Santangelo Philip, Nitin Nitin, Bao Gang
Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA.
Ann Biomed Eng. 2006 Jan;34(1):39-50. doi: 10.1007/s10439-005-9003-6. Epub 2006 Feb 7.
The ability to visualize in real-time the expression level and localization of specific RNAs in living cells can offer tremendous opportunities for biological and disease studies. Here we review the recent development of nanostructured oligonucleotide probes for living cell RNA detection, and discuss the biological and engineering issues and challenges of quantifying gene expression in vivo. In particular, we describe methods that use dual FRET (fluorescence resonance energy transfer) or single molecular beacons in combination with peptide-based or membrane-permeabilization-based delivery, to image the relative level, localization, and dynamics of RNA in live cells. Examples of detecting endogenous mRNAs, as well as imaging their subcellular localization and colocalization are given to illustrate the biological applications, and issues in molecular beacon design, probe delivery, and target accessibility are discussed. The nanostructured probes promise to open new and exciting opportunities in sensitive gene detection for a wide range of biological and medical applications.
实时可视化活细胞中特定RNA的表达水平和定位的能力可为生物学和疾病研究提供巨大机遇。在此，我们综述了用于活细胞RNA检测的纳米结构寡核苷酸探针的最新进展，并讨论了体内基因表达定量中的生物学和工程问题及挑战。特别地，我们描述了使用双荧光共振能量转移（FRET）或单分子信标结合基于肽或基于膜通透化的递送方法，以对活细胞中RNA的相对水平、定位和动态进行成像的方法。给出了检测内源性mRNA以及对其亚细胞定位和共定位进行成像的实例来说明生物学应用，并讨论了分子信标设计、探针递送和靶标可及性方面的问题。这些纳米结构探针有望在广泛的生物学和医学应用的灵敏基因检测中开启新的令人兴奋的机遇。
