RNA-based stable isotope probing (RNA-SIP) to unravel intestinal host-microbe interactions

Methods. 2018 Oct 1:149:25-30. doi: 10.1016/j.ymeth.2018.05.022. Epub 2018 May 30.

Abstract

The RNA-SIP technology, introduced into molecular microbial ecology in 2002, is an elegant technique to link the structure and function of complex microbial communities, i.e. to identify microbial key-players involved in distinct degradation and assimilation processes under in-situ conditions. Due to its dependence of microbial RNA, this technique is particularly suited for environments with high numbers of very active, i.e. significantly RNA-expressing, bacteria. So far, it was mainly used in environmental studies using microbiotas from soil or water habitats. Here we outline and summarize our application of RNA-SIP for the identification of bacteria involved in the degradation and assimilation of prebiotic carbohydrates in intestinal samples of human and animal origin. Following an isotope label from a prebiotic substrate into the RNA of distinct bacterial taxa will help to better understand the functionality of these medically and economically important nutrients in an intestinal environment.

Keywords: Digestion; Nutrition; Prebiotics; RNA-SIP; Substrate utilization.

Publication types

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

MeSH terms

  • Gastrointestinal Microbiome / physiology*
  • Host Microbial Interactions / physiology*
  • Humans
  • Isotope Labeling / instrumentation
  • Isotope Labeling / methods*
  • Microbiota / physiology*
  • RNA Probes / analysis
  • RNA Probes / metabolism*
  • Ultracentrifugation / methods

Substances

  • RNA Probes