NOV 10, 2016 10:00 AM PST

How Do You effectively Detect Heterogeneity in Single-cells?

Presented at: Microbiology
Abstract

Bulk analysis often leads to conclusions that assume averages reflect the dominant biological mechanism operating within an entire population. To fully understand how cellular heterogeneity contributes to biological function, a single-cell analysis approach must be applied.
 
Join us for short talks on breakthroughs in single-cell gene expression research that have advanced our understanding of the immune response and are paving the way towards unmasking gene expression heterogeneity
 
Mechanisms of joint inflammation associated with Chikungunya infection
Dr. June Ann D’Angelo

 
Chikungunya virus (CHIKV) an Old World alphavirus whose primary disease manifestation in humans is fever accompanied by debilitating joint inflammation. Locally produced virus particles disseminate via the circulatory system to different organs including joints. For most, the symptoms clear in a week, however ~10-20% have joint pain that persists for months and is poorly managed with current treatments. We will discuss the use of single-cell gene expression analysis to better understand the immunopathological mechanism that underlies persistent joint inflammation in order to develop safe and effective medicinal interventions. Our research will investigate the hypothesis that CHIKV-associated joint inflammation and pathology is driven by the infection and maintenance of immune cells within the joint.

Single-cell analysis of virus infection:  Zika and beyond    
Eric Clambey

 
The heterogeneity of RNA expression at the single-cell level is often obscured by conventional assays of pooled cellular material. We have recently used the PrimeFlowTM RNA assay to detect virus and host RNAs by flow cytometry, enabling single-cell analysis of virus-host dynamics. We have applied this methodology to detect Zika virus infection, simultaneously quantifying the plus-strand RNA genome and 3’ untranslated region. In parallel, we have used this platform to study herpesvirus-induced host shutoff, directly observing host mRNA degradation in virus-infected cells. These studies have revealed unanticipated heterogeneity in virus-infected cells, and identified new inter-relationships between virus and host RNAs at the single cell level.


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