IL-10 CRISPR Activation Plasmid (m): sc-421076-ACT

Mouse Il10 encodes interleukin-10 (IL-10), a pleiotropic anti-inflammatory cytokine that restrains innate and adaptive immune activation to preserve tissue homeostasis. IL-10 signaling through the IL-10 receptor engages JAK1/TYK2 and STAT3 to suppress pro-inflammatory cytokine production, limit antigen-presenting cell maturation, and modulate T cell polarization and regulatory programs. This axis is central to mucosal immune tolerance and resolution of inflammation, with relevance to immune-mediated pathology such as colitis-like inflammation, chronic infection–associated immunoregulation, and tumor microenvironment immune suppression in experimental models. Altered IL-10 activity is therefore widely studied in macrophages, dendritic cells, B cells, and T cells to understand cytokine networks and inflammatory signaling circuits.

IL-10 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Il10 expression without altering the underlying DNA sequence.

IL-10 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Il10 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 Il10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IL-10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Il10 locus and enabling the study of IL-10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IL-10 pathway restoration in tumor cells with silenced or reduced Il10 expression.

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