IL-6 CRISPR Activation Plasmid (h): sc-400390-ACT

Human IL6 encodes interleukin-6 (IL-6), a pleiotropic cytokine that coordinates acute-phase responses, innate and adaptive immune crosstalk, and hematopoietic regulation. IL-6 signals through IL6R and the gp130 co-receptor (IL6ST) to activate JAK/STAT3, MAPK/ERK, and PI3K/AKT pathways, shaping transcriptional programs linked to inflammation, survival, and differentiation. Dysregulated IL-6 activity is implicated in chronic inflammatory states, autoimmunity, and tumor-associated inflammation, where it can influence myeloid polarization, T helper lineage balance, and angiogenic or metabolic remodeling. As a central node in cytokine networks, IL6 is frequently studied for its role in feedback regulation of NF-κB-driven responses and stress-induced transcriptional circuits.

IL-6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IL6 expression without altering the underlying DNA sequence.

IL-6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IL6 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 IL6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IL-6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IL6 locus and enabling the study of IL-6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IL-6 pathway restoration in tumor cells with silenced or reduced IL6 expression.

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