Live cell gene expression detection in monocytes and macrophages through flow cytometry (TECH1P.837)

• Published in: The Journal of immunology (1950) Vol. 192; no. 1_Supplement; pp. 69 - 69.5
• Main Authors: Weldon, Don, Williams, Yuko, Liu, Haizhen, Ko, Alex, Koong, Victor
• Format: Journal Article
• Language: English
• Published: 01.05.2014
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.192.Supp.69.5

Abstract Detecting gene expression in immune cells has traditionally been limited to technologies that utilize a reporter construct for a gene of interest or within lysed or fixed cell populations as is the case for RT-PCR. Here we show the ability to detect gene expression in live monocytes and macrophages which allows for more physiologically relevant information based on the cell’s response to given stimuli. Determining which genes were up or down regulated in those cells provides insight into complex gene regulatory networks and immune cell function. Here we present a novel RNA expression detection technology capable of detecting specific mRNA and miRNA in live, intact cells. This technology allows for detection of target RNA, with the ability to perform downstream assays such as Fluorescence-Activated Cell Sorting (FACS) to enrich for a specific subpopulation of live cells based solely on their RNA expression profile. Following FACS sorting based on RNA detection the cells remain viable and fully functional for use in downstream assays allowing researchers the ability to further characterize or determine functionality of the enriched population. Within a population of cells there is inherent homogeneity which can skew results when only looking at the population level as opposed to understanding the single cell resolution of gene expression. Looking at RNA levels in a more dynamic manner proves to be much more informative than looking at duplicate samples over time.