What have single-molecule studies taught us about gene expression?
The production of a single mRNA is the result of many sequential steps, from docking of transcription factors to polymerase initiation, elongation, splicing, and, finally, termination. Much of our knowledge about the fundamentals of RNA synthesis and processing come from ensemble in vitro biochemical measurements. Single-molecule approaches are very much in this same reductionist tradition but offer exquisite sensitivity in space and time along with the ability to observe heterogeneous behavior and actually manipulate macromolecules. These techniques can also be applied in vivo, allowing one to address questions in living cells that were previously restricted to reconstituted systems. In this review, we examine the unique insights that single-molecule techniques have yielded on the mechanisms of gene expression.
Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Huimin
H
Chen
Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Daniel R
0000-0001-9253-3055
DR
Larson
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005786:Gene Expression Regulation; D005821:Genetic Techniques / Q000592:standards / Q000639:trends; D006801:Humans; D010441:Peptide Chain Elongation, Translational; D012326:RNA Splicing; D014157:Transcription Factors / Q000378:metabolism
D014157:Transcription Factors; D012321:DNA-Directed RNA Polymerases
fluorescence; single molecule; splicing; transcription