核糖核酸
抄写(语言学)
点击化学
尿苷
环加成
体内
化学
核酸
信使核糖核酸
生物化学
分子生物学
生物
细胞生物学
催化作用
组合化学
基因
遗传学
哲学
语言学
作者
Cindy Y. Jao,Adrian Salic
标识
DOI:10.1073/pnas.0808480105
摘要
We describe a chemical method to detect RNA synthesis in cells, based on the biosynthetic incorporation of the uridine analog 5-ethynyluridine (EU) into newly transcribed RNA, on average once every 35 uridine residues in total RNA. EU-labeled cellular RNA is detected quickly and with high sensitivity by using a copper (I)-catalyzed cycloaddition reaction (often referred to as "click" chemistry) with fluorescent azides, followed by microscopic imaging. We demonstrate the use of this method in cultured cells, in which we examine the turnover of bulk RNA after EU pulses of varying lengths. We also use EU to assay transcription rates of various tissues in whole animals, both on sections and by whole-mount staining. We find that total transcription rates vary greatly among different tissues and among different cell types within organs.
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