Characterization of Gene Repression by Designed Transcription Activator-like Effector Dimer Proteins

Nicole A. Becker, Justin P. Peters, Tanya L. Schwab, William J. Phillips, Jordan P. Wallace, Karl J. Clark, L. James Maher

Research output: Contribution to journalArticlepeer-review

Abstract

Gene regulation by control of transcription initiation is a fundamental property of living cells. Much of our understanding of gene repression originated from studies of the Escherichia coli lac operon switch, in which DNA looping plays an essential role. To validate and generalize principles from lac for practical applications, we previously described artificial DNA looping driven by designed transcription activator-like effector dimer (TALED) proteins. Because TALE monomers bind the idealized symmetrical lac operator sequence in two orientations, our prior studies detected repression due to multiple DNA loops. We now quantitatively characterize gene repression in living E. coli by a collection of individual TALED loops with systematic loop length variation. Fitting of a thermodynamic model allows unequivocal demonstration of looping and comparison of the engineered TALED repression system with the natural lac repressor system.

Original languageEnglish (US)
Pages (from-to)2045-2054
Number of pages10
JournalBiophysical Journal
Volume119
Issue number10
DOIs
StatePublished - Nov 17 2020

ASJC Scopus subject areas

  • Biophysics

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