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Updated: May 15, 2026

Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages
Published on: November 3, 2014
Small Interfering RNA (siRNA)-Mediated Knockdown of Macrophage Migration Inhibitory Factor Transcript in Adherent and
Swagata Ghosh1, Mathan L2, Hari Vignesh Sekar3
1Department of Microbiology, Aravind Medical Research Foundation, Madurai, India. swagata@aravind.org.
Abstract:
Small interfering RNA (siRNA)-mediated depletion of macrophage migration inhibitory factor (MIF) in cell culture systems is essential for understanding the function or relevance of MIF in various cell types under different experimental conditions. In this article, we provide a detailed description of the methods for siRNA transfection in a terminally differentiated adherent cell line derived from human corneal epithelial cells. Furthermore, we discuss siRNA transfection techniques for two suspension cell lines of monocytic and promyelocytic origin, which can be differentiated into adherent macrophage lines and suspension neutrophil lines, respectively. Following siRNA transfection, we evaluated MIF knockdown at the transcript level through RNA extraction, cDNA synthesis, and real-time PCR analysis.
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