DOI: 10.1101/476366Nov 22, 2018Paper

Interrogation of genes controlling biofilm formation using CRISPR interference in Pseudomonas fluorescens

BioRxiv : the Preprint Server for Biology
Marie-Francoise Noirot-GrosPhilippe H Noirot

Abstract

Bacterial biofilm formation involves multigenic signaling and regulatory pathways that control the transition from motile to sessile lifestyle, production of extracellular polymeric matrix, and maturation of the biofilm complex 3D structure. Biofilms are extensively studied because of their importance in biomedical, ecological and industrial settings. Genetic approaches based on gene inactivation are powerful for mechanistic studies but often are labor intensive, limiting systematic gene surveys to the most tractable bacterial hosts. Here, we adapted the CRISPR interference (CRISPRi) system for use in P. fluorescens. We found that CRISPRi is applicable to three genetically and physiologically diverse species, SBW25, WH6 and Pf0-1 and affords extended periods of time to study complex phenotypes such as cell morphology, motility and biofilm formation. In SBW25, CRISPRi-mediated silencing of the GacA/S two-component system and genes regulated by cylic-di-GMP produced phenotypes similar to those previously described after gene inactivation in various Pseudomonas. Combined with detailed confocal microscopy of biofilms, our study also revealed novel phenotypes associated with biofilm architecture and extracellular matrix biosynthesis...Continue Reading

Related Concepts

Cell Motility
Classification
Extracellular Matrix
Genes
Genetic Techniques
Guanosine Monophosphate
Pseudomonas
Pseudomonas fluorescens
Microbial Biofilms
Microscopy, Confocal

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