MIF CRISPR Activation Plasmid (h): sc-400574-ACT

Macrophage migration inhibitory factor (MIF) is a pleiotropic human cytokine with tautomerase activity that regulates innate and adaptive immune responses, cell survival, and stress signaling. It functions through receptors including CD74 with co-receptors such as CXCR2/CXCR4, engaging downstream MAPK/ERK, PI3K/AKT, and NF-κB pathways to shape inflammatory gene expression and leukocyte recruitment. MIF also intersects with glucocorticoid signaling and redox homeostasis, influencing macrophage polarization, angiogenic programs, and tumor–immune microenvironment dynamics. Dysregulated MIF expression has been associated with chronic inflammatory disorders, fibrosis, and cancer-related processes, making it a useful node for mechanistic studies of immune regulation and disease-relevant signaling networks.

MIF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MIF expression without altering the underlying DNA sequence.

MIF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MIF locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the MIF transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MIF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MIF locus and enabling the study of MIF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MIF pathway restoration in tumor cells with silenced or reduced MIF expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.