Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, VIC, Australia.
Department of Molecular and Translational Science, Monash University, Clayton, VIC, Australia.
Front Cell Infect Microbiol. 2018 Mar 19;8:87. doi: 10.3389/fcimb.2018.00087. eCollection 2018.
Murine models of serovar Typhimurium infection are one of the commonest tools to study host-pathogen interactions during bacterial infections. Critically, the outcome of . 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 . 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 . 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.
鼠伤寒沙门氏菌感染的鼠模型是研究细菌感染过程中宿主-病原体相互作用的常用工具之一。关键的是，用于感染的小鼠品系的遗传背景会影响鼠伤寒沙门氏菌感染的结果，来自 C57BL/6 和 BALB/c 小鼠的巨噬细胞缺乏控制细胞内细菌复制的能力。出于这个原因，使用同源性导入系，将天然保护小鼠品系的遗传背景与易感品系的遗传背景混合，有可能显著改变鼠伤寒沙门氏菌感染研究的结果和解释。在这里，我们描述了如何使用 CRISPR/Cas9 基因编辑生成巨噬细胞敲除细胞系，以帮助确定同源性导入系的初级巨噬细胞中背景污染对鼠伤寒沙门氏菌感染研究结果的表型的贡献。我们自己的经验说明了 CRISPR/Cas9 技术如何可以补充现有的敲除模型，并表明同时进行两种遗传模型的研究具有很大的价值，可以排除对宿主-病原体相互作用的意外副作用。
