A modular degron library for synthetic circuits in mammalian cells

Nat Commun. 2019 May 1;10(1):2013. doi: 10.1038/s41467-019-09974-5.

Abstract

Tight control over protein degradation is a fundamental requirement for cells to respond rapidly to various stimuli and adapt to a fluctuating environment. Here we develop a versatile, easy-to-handle library of destabilizing tags (degrons) for the precise regulation of protein expression profiles in mammalian cells by modulating target protein half-lives in a predictable manner. Using the well-established tetracycline gene-regulation system as a model, we show that the dynamics of protein expression can be tuned by fusing appropriate degron tags to gene regulators. Next, we apply this degron library to tune a synthetic pulse-generating circuit in mammalian cells. With this toolbox we establish a set of pulse generators with tailored pulse lengths and magnitudes of protein expression. This methodology will prove useful in the functional roles of essential proteins, fine-tuning of gene-expression systems, and enabling a higher complexity in the design of synthetic biological systems in mammalian cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Amino Acid Sequence / genetics*
  • Biotechnology / methods
  • Gene Expression Regulation*
  • HEK293 Cells
  • Half-Life
  • HeLa Cells
  • Humans
  • Intravital Microscopy / methods
  • Mesenchymal Stem Cells
  • Microscopy, Fluorescence
  • Protein Engineering / methods*
  • Proteolysis*
  • Synthetic Biology / methods