Pederson T
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 377 Plantation Street, Worcester, MA 01605, USA.
Nucleic Acids Res. 2001 Mar 1;29(5):1013-6. doi: 10.1093/nar/29.5.1013.
The advent of jellyfish green fluorescent protein and its spectral variants, together with promising new fluorescent proteins from other classes of the Cnidarian phylum (coral and anemones), has greatly enhanced and promises to further boost the detection and localization of proteins in cell biology. It has been less widely appreciated that highly sensitive methods have also recently been developed for detecting the movement and localization in living cells of the very molecules that precede proteins in the gene expression pathway, i.e. RNAs. These approaches include the microinjection of fluorescent RNAs into living cells, the in vivo hybridization of fluorescent oligonucleotides to endogenous RNAs and the expression in cells of fluorescent RNA-binding proteins. This new field of 'fluorescent RNA cytochemistry' is summarized in this article, with emphasis on the biological insights it has already provided. These new techniques are likely to soon collaborate with other emerging approaches to advance the investigation of RNA birth, RNA-protein assembly and ribonucleoprotein particle transport in systems such as oocytes, embryos, neurons and other somatic cells, and may even permit the observation of viral replication and transcription pathways as they proceed in living cells, ushering in a new era of nucleic acids research in vivo.
水母绿色荧光蛋白及其光谱变体的出现,以及来自刺胞动物门其他类群(珊瑚和海葵)的前景广阔的新型荧光蛋白,极大地增强了细胞生物学中蛋白质检测和定位的能力,而且有望进一步提升。人们尚未广泛认识到,最近还开发出了高灵敏度方法,用于检测基因表达途径中先于蛋白质的分子,即RNA,在活细胞中的移动和定位。这些方法包括将荧光RNA显微注射到活细胞中、荧光寡核苷酸与内源性RNA的体内杂交,以及荧光RNA结合蛋白在细胞中的表达。本文总结了“荧光RNA细胞化学”这一新领域,重点介绍了它已提供的生物学见解。这些新技术可能很快会与其他新兴方法合作,推动对卵母细胞、胚胎、神经元和其他体细胞等系统中RNA的产生、RNA-蛋白质组装以及核糖核蛋白颗粒运输的研究,甚至可能允许观察病毒在活细胞中的复制和转录途径,开创体内核酸研究的新时代。