Building a Synthetic Transcriptional Oscillator

Methods Mol Biol. 2016:1342:185-99. doi: 10.1007/978-1-4939-2957-3_10.

Abstract

Reaction circuits mimicking genetic oscillators can be realized with synthetic, switchable DNA genes (so-called genelets), and two enzymes only, an RNA polymerase and a ribonuclease. The oscillatory behavior of the genelets is driven by the periodic production and degradation of RNA effector molecules. Here, we describe the preparation, assembly, and testing of a synthetic, transcriptional two-node negative-feedback oscillator, whose dynamics can be followed in real-time by fluorescence read-out.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Feedback, Physiological
  • Fluorometry
  • Genes, Synthetic / genetics*
  • Genetic Engineering / methods*
  • RNA, Messenger / genetics
  • RNA, Messenger / isolation & purification
  • RNA, Messenger / metabolism
  • Transcription, Genetic / genetics*

Substances

  • RNA, Messenger
  • DNA