Skip to Content
Merck
  • A Synthetic Mammalian Therapeutic Gene Circuit for Sensing and Suppressing Inflammation.

A Synthetic Mammalian Therapeutic Gene Circuit for Sensing and Suppressing Inflammation.

Molecular therapy : the journal of the American Society of Gene Therapy (2017-01-28)
Anže Smole, Duško Lainšček, Urban Bezeljak, Simon Horvat, Roman Jerala
ABSTRACT

Inflammation, which is a highly regulated host response against danger signals, may be harmful if it is excessive and deregulated. Ideally, anti-inflammatory therapy should autonomously commence as soon as possible after the onset of inflammation, should be controllable by a physician, and should not systemically block beneficial immune response in the long term. We describe a genetically encoded anti-inflammatory mammalian cell device based on a modular engineered genetic circuit comprising a sensor, an amplifier, a "thresholder" to restrict activation of a positive-feedback loop, a combination of advanced clinically used biopharmaceutical proteins, and orthogonal regulatory elements that linked modules into the functional device. This genetic circuit was autonomously activated by inflammatory signals, including endogenous cecal ligation and puncture (CLP)-induced inflammation in mice and serum from a systemic juvenile idiopathic arthritis (sIJA) patient, and could be reset externally by a chemical signal. The microencapsulated anti-inflammatory device significantly reduced the pathology in dextran sodium sulfate (DSS)-induced acute murine colitis, demonstrating a synthetic immunological approach for autonomous anti-inflammatory therapy.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
1,2-Dilinoleoyl-3-palmitoyl-rac-glycerol, ≥95% (TLC), liquid
Sigma-Aldrich
Formalin solution, neutral buffered, 10%, histological tissue fixative
Sigma-Aldrich
Mayer′s Hematoxylin Solution
Sigma-Aldrich
Poly-L-lysine hydrobromide, mol wt 15,000-30,000 by viscosity
Sigma-Aldrich
IL-1Ra human, recombinant, expressed in E. coli, ≥98% (SDS-PAGE), ≥98% (HPLC), suitable for cell culture