MICB CRISPR Activation Plasmid (h): sc-401524-ACT

MICB encodes MHC class I polypeptide-related sequence B, a stress-inducible ligand displayed on the surface of human cells that engages the activating receptor NKG2D on NK cells and subsets of T cells. MICB expression is regulated by cellular stress programs including DNA damage responses, oxidative stress, and infection-associated signaling, linking innate immune surveillance to changes in proteostasis and cell cycle control. As part of the NKG2D ligand axis, MICB participates in immunorecognition pathways that influence cytotoxic lymphocyte activation and cytokine production. Dysregulated MICB expression or shedding has been associated with immune evasion phenotypes and inflammatory signaling in cancer biology and chronic infection models, supporting its use in studies of tumor–immune interactions.

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

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

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