LEREPO4 CRISPR Activation Plasmid (m): sc-427290-ACT

Mouse Zc3h15 encodes the LEREPO4 protein, a zinc finger CCCH-type RNA-binding factor implicated in post-transcriptional gene regulation, including mRNA stability and translation control. By modulating RNA processing and turnover, Zc3h15 can influence signaling outputs downstream of innate immune and stress-response programs that depend on rapid transcript remodeling. Altered activity of CCCH zinc finger RNA-binding proteins is frequently linked to dysregulated cytokine expression, cellular stress adaptation, and aberrant proliferation, making Zc3h15 a useful node for mechanistic studies of RNA-centric regulation. In biomedical research models, perturbing Zc3h15 supports pathway dissection in inflammation, hematopoietic regulation, and other contexts where RNA fate decisions shape phenotype.

LEREPO4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Zc3h15 expression without altering the underlying DNA sequence.

LEREPO4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Zc3h15 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 Zc3h15 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LEREPO4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Zc3h15 locus and enabling the study of LEREPO4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LEREPO4 pathway restoration in tumor cells with silenced or reduced Zc3h15 expression.

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