TTP CRISPR Activation Plasmid (h): sc-400722-ACT

Human ZFP36 encodes tristetraprolin (TTP), an RNA-binding protein that recognizes AU-rich elements in the 3′ UTR of target transcripts and promotes their deadenylation and decay. Through post-transcriptional control, TTP constrains cytokine and immediate-early gene expression and shapes inflammatory and stress-response programs, including TNF/NF-κB-driven signaling. TTP activity is dynamically regulated by phosphorylation downstream of MAPK pathways, influencing mRNA turnover, translation, and ribonucleoprotein granule dynamics. Dysregulated ZFP36/TTP expression or function has been associated with aberrant immune activation, chronic inflammatory phenotypes, and altered tumor-associated cytokine networks, making it a useful node for mechanistic studies of inflammation-linked disease biology.

TTP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZFP36 expression without altering the underlying DNA sequence.

TTP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZFP36 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 ZFP36 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TTP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZFP36 locus and enabling the study of TTP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TTP pathway restoration in tumor cells with silenced or reduced ZFP36 expression.

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