MCPIP CRISPR Activation Plasmid (h): sc-401790-ACT

ZC3H12A encodes MCPIP (also known as Regnase-1), an RNA-binding endoribonuclease that restrains inflammatory gene expression by promoting decay of cytokine and chemokine mRNAs and modulating microRNA biogenesis. MCPIP integrates signals downstream of innate immune receptors and cytokine pathways, including NF-κB and MAPK cascades, to shape macrophage activation, T cell responses, and resolution of inflammation. Through regulation of transcript stability and immune cell differentiation programs, ZC3H12A is implicated in dysregulated immune homeostasis observed in autoimmune and chronic inflammatory disease contexts. Its activity also intersects with cellular stress responses and apoptosis-related signaling, making it relevant for studies of inflammation-linked tissue damage and tumor–immune interactions.

MCPIP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZC3H12A expression without altering the underlying DNA sequence.

MCPIP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZC3H12A 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 ZC3H12A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MCPIP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZC3H12A locus and enabling the study of MCPIP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MCPIP pathway restoration in tumor cells with silenced or reduced ZC3H12A expression.

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