MS4A6A CRISPR Activation Plasmid (h): sc-417401-ACT

MS4A6A encodes a member of the membrane-spanning 4A (MS4A) family, a group of tetraspan-like proteins enriched in hematopoietic and myeloid lineages. MS4A6A is frequently used as a marker of microglia and macrophage activation states and is implicated in membrane organization and regulation of cell-surface signaling complexes that shape innate immune responses. Expression of MS4A6A tracks with inflammatory programs and pathways linked to immune cell activation, phagocytic function, and cytokine-driven transcriptional states. Altered MS4A6A expression has been associated with neuroinflammation-related phenotypes and is commonly studied in the context of neurodegenerative disease genetics and microglial biology.

MS4A6A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MS4A6A expression without altering the underlying DNA sequence.

MS4A6A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MS4A6A 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 MS4A6A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MS4A6A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MS4A6A locus and enabling the study of MS4A6A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MS4A6A pathway restoration in tumor cells with silenced or reduced MS4A6A expression.

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