The Use of CRISPR/Cas9 Gene Editing to Confirm Congenic Contaminations in Host-Pathogen Interaction Studies

Frontiers in Cellular and Infection Microbiology

Wu, Di¹  Ferrand, Jonathan²  pin, Geneviè²  Pedersen, John²  Pé³  Croft, Nathan P.⁴  ve⁴  Behlke, Mark A.⁴  Mangan, Niamh E.⁵  Diener, Kerrilyn R.⁶  Purcell, Anthony W.⁷  Hayball, John D.⁸ 

[1] Adelaide Medical School, Robinson Research Institute, The University of Adelaide, Australia;Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Australia;Department of Biochemistry and Molecular Biology, Infection and Immunity Program, Biomedicine Discovery Institute, Monash University, Australia;Department of Molecular and Translational Science, Monash University, Australia;Department of Periodontology, University of North Carolina at Chapel Hill, United States;Experimental Therapeutics Laboratory, School of Pharmacy and Medical Science, Sansom Institute for Health Research, University of South Australia, Australia;Integrated DNA Technologies Inc., United States;TissuPath Specialist Pathology, Australia

关键词: Salmonella;    CRISPR/Cas9;    congenic mice;    Background contamination;    Host-Pathogen Interactions;   

DOI  :  10.3389/fcimb.2018.00087

学科分类：生物科学（综合）

来源: Frontiers

【 摘 要 】

Murine models of Salmonella enterica serovar Typhimurium infection are one of the commonest tools to study host-pathogen interactions during bacterial infections. Critically, the outcome of S. Typhimurium infection is impacted by the genetic background of the mouse strain used, with macrophages from C57BL/6 and BALB/c mice lacking the capacity to control intracellular bacterial replication. For this reason, the use of congenic strains, which mix the genetic backgrounds of naturally protected mouse strains with those of susceptible strains, has the capacity to significantly alter results and interpretation of S. Typhimurium infection studies. Here, we describe how macrophage knockout cell lines generated by CRISPR/Cas9 gene editing can help determine the contribution of background contaminations in the phenotypes of primary macrophages from congenic mice, on the outcome of S. Typhimurium infection studies. Our own experience illustrates how the CRISPR/Cas9 technology can be used to complement pre-existing knockout models, and shows that there is great merit in performing concurrent studies with both genetic models, to exclude unanticipated side-effects on host-pathogen interactions.

【 授权许可】

CC BY