title: Gene Expression Profiling of Infecting Microbes Using a Digital Bar-coding Platform.
abstract: For most mammalian pathogens, gene expression profiling studies have been limited by technical difficulties to accurately quantify pathogen gene transcripts from infected tissues. Pathogen RNA constitutes a tiny portion of the total RNA isolated from infected tissue samples. Both microarray and RNAseq technologies have difficulties in generating reliable reads for weakly expressed pathogen genes. Mutant pathogen strains with reduced in vivo proliferation pose an even bigger challenge. Here we describe an in vivo gene expression profiling protocol that is very fast, extremely sensitive and highly reproducible. We developed this protocol during our investigation of the fungal pathogen Candida albicans in a murine model of hematogenously disseminated candidiasis. Using this protocol, we have documented time courses of dynamically regulated C. albicans gene expression during kidney infection, and discovered unexpected features of gene expression responses to antifungal drug treatment in vivo.
authors: Department of Biological Sciences, Carnegie Mellon University; wxu7206@gmail.com.
authors: Wenjie
authors: W
authors: Xu
authors: Infectious Diseases, Harbor-UCLA Medical Center.
authors: Norma V
authors: NV
authors: Solis
authors: Infectious Diseases, Harbor-UCLA Medical Center.
authors: Scott G
authors: SG
authors: Filler
authors: Department of Biological Sciences, Carnegie Mellon University.
authors: Aaron P
authors: AP
authors: Mitchell
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D002177:Candidiasis / Q000188:drug therapy / Q000235:genetics*; D004195:Disease Models, Animal; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling / Q000379:methods*; D051379:Mice; D046228:Microarray Analysis; D012313:RNA / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D012313:RNA