MICA CRISPR Activation Plasmid (h): sc-418864-ACT

MICA (MHC class I polypeptide-related sequence A) encodes a stress-inducible cell-surface glycoprotein that functions as a ligand for the activating receptor NKG2D on NK cells and subsets of CD8+ T cells. By engaging NKG2D, MICA contributes to immune surveillance pathways that sense cellular stress, infection, and transformation, influencing cytotoxic activation and cytokine signaling at the immunological synapse. MICA expression is shaped by DNA damage responses, heat shock pathways, and inflammatory signaling, and its shedding or altered surface presentation can modulate receptor engagement. Dysregulated MICA–NKG2D signaling has been implicated in tumor immune evasion, chronic inflammatory states, and autoimmune disease-associated immune activation.

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

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

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