Tollip CRISPR Activation Plasmid (h): sc-402638-ACT

Human TOLLIP encodes Toll-interacting protein (Tollip), an endosomal adaptor that modulates innate immune signaling downstream of Toll-like receptors and the IL-1 receptor family. Tollip binds ubiquitinated cargo and interfaces with the ubiquitin–proteasome and endolysosomal trafficking machinery to influence receptor turnover, signal amplitude, and inflammatory gene expression. Through its roles in TLR/IL-1R–NF-κB and MAPK pathway regulation, TOLLIP contributes to cellular decisions that shape cytokine production and immune tolerance. Dysregulated TOLLIP activity has been associated with altered inflammatory responses and has been investigated in contexts including infection biology, chronic inflammation, and immune-mediated disease mechanisms.

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

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

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