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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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Small Interfering RNA (siRNA)-Mediated Knockdown of Macrophage Migration Inhibitory Factor Transcript in Adherent and Suspension Cultures of Human Cell Lines

  • 实验方案

  • 发表日期：

    2026年

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  • Swagata Ghosh,
  • Mathan L,
  • Hari Vignesh Sekar

摘要

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.