Single molecule studies of RNA polymerase II transcription in vitro

Abstract

Eukaryotic mRNA transcription by RNA polymerase II (RNAP II) is the first step in gene expression and a key determinant of cellular regulation. Elucidating the mechanism by which RNAP II synthesizes RNA is therefore vital to determining how genes are controlled under diverse biological conditions. Significant advances in understanding RNAP II transcription have been achieved using classical biochemical and structural techniques; however, aspects of the transcription mechanism cannot be assessed using these approaches. The application of single-molecule techniques to study RNAP II transcription has provided new insight only obtainable by studying molecules in this complex system one at a time.

Keywords: :

• transcription
• RNA polymerase II
• single-molecule
• optical tweezers
• fluorescence
• FRET
• TIRF

Disclosure of Potential Conflicts of Interest

No potential conflicts of interest were disclosed.

Acknowledgments

This work was funded by grant MCB-1244518 from the National Science Foundation. A.E.H. was supported in part by NIH Training Grant T32 GM-065103.