Direct Observation of Single RNA Polymerase Processing through a Single Endogenous Gene In a Living Yeast Cell

摘要

The transcription of a gene into messenger RNA (mRNA) by the RNA polymerase II (RNAPII) is the first, fundamental step in gene expression and consists of the three major phases initiation, elongation, and termination. Each step of transcription is highly regulated by the binding and action of numerous transcription factors, and functional understanding of transcriptional networks and their dynamics is crucial for the understanding of cellular systems and their development. Classical biochemical studies of transcription are based on the determination of expression levels by the isolation of mRNA from cell populations. The applied bulk techniques, such as northern blotting and reverse-transcription quantitative PCR, however, comprehend the drawback of averaging over a large number of cells and hence obscure cell-to-cell variation and lack spatial information. Also, the dynamics and regulation of single transcription events cannot be resolved.