Skint8 CRISPR Activation Plasmid (m2): sc-437039-ACT-2

Mouse Skint8 (selection and upkeep of intraepithelial T cells protein 8) is a member of the SKINT family, an immunoglobulin superfamily group implicated in epithelial–immune communication and the regulation of tissue-resident lymphocyte phenotypes. Skint8 expression is linked to immune homeostasis at barrier surfaces, potentially influencing antigen recognition contexts, cytokine signaling networks, and differentiation or maintenance programs of specialized T cell subsets within the epithelium. Dysregulation of SKINT-family pathways is of interest for studies of cutaneous and mucosal inflammation, host defense, and immune dysregulation mechanisms relevant to autoimmunity and tumor immune surveillance. Skint8 gene editing or perturbation in mouse models supports functional interrogation of epithelial cell–intrinsic immunoregulatory circuits, cell–cell interaction biology, and pathway mapping in immunology and skin research workflows.

Skint8 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Skint8 expression without altering the underlying DNA sequence.

Skint8 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Skint8 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 Skint8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Skint8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Skint8 locus and enabling the study of Skint8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Skint8 pathway restoration in tumor cells with silenced or reduced Skint8 expression.

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