TNFα-IP 2 CRISPR Activation Plasmid (h): sc-401685-ACT

TNFAIP2 (TNFα-induced protein 2; TNFα-IP 2) is an inflammation-responsive gene induced by TNFα and other NF-κB–activating stimuli, linking cytokine signaling to cellular remodeling programs. The protein has been implicated in regulation of actin cytoskeleton dynamics, membrane trafficking, and cell motility, supporting processes such as adhesion and migration in immune and stromal contexts. TNFAIP2 expression tracks with activated inflammatory states and is frequently studied in pathways connecting innate immune signaling, vascular/endothelial activation, and tissue remodeling. Dysregulated TNFAIP2 has been associated with pathological inflammation and cancer-related phenotypes, motivating mechanistic studies of how NF-κB-dependent transcriptional outputs shape cellular behavior.

TNFα-IP 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNFAIP2 expression without altering the underlying DNA sequence.

TNFα-IP 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNFAIP2 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 TNFAIP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TNFα-IP 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNFAIP2 locus and enabling the study of TNFα-IP 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TNFα-IP 2 pathway restoration in tumor cells with silenced or reduced TNFAIP2 expression.

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