Tunable expression tools enable single-cell strain distinction in the gut microbiome

• Weston R. Whitaker ^[1] ; Elizabeth Stanley Shepherd ; Justin L. Sonnenburg
  1. [1] Novome Biotechnologies
• Localización: Cell, ISSN 0092-8674, Vol. 169, Nº. 3, 2017, págs. 538-546
• Idioma: inglés
• Texto completo no disponible (Saber más ...)
• Resumen

  • Applying synthetic biology to engineer gut-resident microbes provides new avenues to investigate microbe-host interactions, perform diagnostics, and deliver therapeutics. Here, we describe a platform for engineering Bacteroides, the most abundant genus in the Western microbiota, which includes a process for high-throughput strain modification. We have identified a novel phage promoter and translational tuning strategy and achieved an unprecedented level of expression that enables imaging of fluorescent-protein-expressing Bacteroides stably colonizing the mouse gut. A detailed characterization of the phage promoter has provided a set of constitutive promoters that span over four logs of strength without detectable fitness burden within the gut over 14 days. These promoters function predictably over a 1,000,000-fold expression range in phylogenetically diverse Bacteroides species. With these promoters, unique fluorescent signatures were encoded to allow differentiation of six species within the gut. Fluorescent protein-based differentiation of isogenic strains revealed that priority of gut colonization determines colonic crypt occupancy.