TY - JOUR T1 - Expression Microdissection for use in qPCR based analysis of miRNA in a single cell type JF - bioRxiv DO - 10.1101/2022.06.24.497524 SP - 2022.06.24.497524 AU - Ana E. Jenike AU - Brady Bunkelman AU - Kira A. Perzel Mandell AU - Cliff Oduor AU - Deborah Chin AU - Devin Mair AU - Katharine M. Jenike AU - Deok-Ho Kim AU - Jeffrey A. Bailey AU - Miriam H. Rafailovich AU - Avi Z. Rosenberg AU - Marc K. Halushka Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/06/28/2022.06.24.497524.abstract N2 - Cell-specific microRNA (miRNA) expression estimates are important in characterizing the localization of miRNA signaling within tissues. Much of this data is obtained from cultured cells, a process known to significantly alter miRNA expression levels. Thus, our knowledge of in vivo cell miRNA expression estimates is poor. We previously demonstrated expression microdissection-miRNA-sequencing (xMD-miRNA-seq) as a means to acquire in vivo estimates, directly from formalin fixed tissues, albeit with limited yield. Here we optimized each step of the xMD process including tissue retrieval, tissue transfer, film preparation, and RNA isolation to increase RNA yields and ultimately show strong enrichment for in vivo miRNA expression by qPCR array. These method improvements, including the development of a non-crosslinked ethylene vinyl acetate (EVA) membrane, resulted in a 23-45 fold increase in miRNA yield, depending on cell type. By qPCR, miR-200a was increased 14-fold in xMD-derived small intestine epithelial cells, with a concurrent 336-fold reduction in miR-143, relative to the matched non-dissected duodenal tissue. xMD is now an optimized method to obtain robust in vivo miRNA expression estimates from cells.Competing Interest StatementThe authors have declared no competing interest. ER -