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Investigation of cell trajectories in chaotic flow fields generated by a helical static micromixer for rapid crosslinking kinetics

20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2016

Priye, Aashish P.; Renzi, Ronald F.; Meagher, Robert M.; Wu, Meiye; Poorey, Kunal N.

Transcription factor (TF) binding pattern in chromatin provide precise and comprehensive information about the cell state. However, current analysis methods such as chromatin immunoprecipitation (ChIP) does not readily facilitate in-vivo characterization required to study short lived chromatin complexes. Here we develop a microfluidic stop flow system which is capable of mixing bacterial cells with formaldehyde on a sub-second mixing residence time scale enabling us to probe the kinetics of TF bound chromatin intermediates.

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TYPE Conference Poster YEAR 2016
ScopusOSTI

A transcriptomic analysis of Yersinia enterocolitica biovar 1B infecting murine macrophages reveals new mechanisms of intracellular survival

Infection and Immunity

Poorey, Kunal N.; Sinha, Anupama S.; Curtis, Deanna J.; Williams, Kelly P.; Branda, Steven B.; Meagher, Robert M.

Yersinia enterocolitica is typically considered an extracellular pathogen; however, during the course of an infection, a significant number of bacteria are stably maintained within host cell vacuoles. Little is known about this population and the role it plays during an infection. To address this question and to elucidate the spatially and temporally dynamic gene expression patterns of Y. enterocoliticabiovar 1B through the course of an in vitro infection, transcriptome sequencing and differential gene expression analysis of bacteria infecting murine macrophage cells were performed under four distinct conditions. Bacteria were first grown in a nutrient-rich medium at 26°C to establish a baseline of gene expression that is unrelated to infection. The transcriptomes of these bacteria were then compared to bacteria grown in a conditioned cell culture medium at 37°C to identify genes that were differentially expressed in response to the increased temperature and medium but not in response to host cells. Infections were then performed, and the transcriptomes of bacteria found on the extracellular surface and intracellular compartments were analyzed individually. The upregulated genes revealed potential roles for a variety of systems in promoting intracellular virulence, including the Ysa type III secretion system, the Yts2 type II secretion system, and the Tad pilus. It was further determined that mutants of each of these systems had decreased virulence while infecting macrophages. Overall, these results reveal the complete set of genes expressed by Y. enterocolitica in response to infection and provide the groundwork for future virulence studies.

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TYPE Journal Article YEAR 2015
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Results 26–47 of 47
Results 26–47 of 47