RoXaN CRISPR Activation Plasmid (h): sc-411701-ACT

ZC3H7B encodes RoXaN, a cytoplasmic RNA-binding protein implicated in post-transcriptional gene regulation through interactions with poly(A)-binding protein and components of messenger ribonucleoprotein complexes. RoXaN has been linked to control of mRNA stability and translation, supporting coordinated expression of genes involved in cellular stress responses and growth-associated programs. Reported interactions with viral factors highlight a role in host–pathogen interface biology and regulation of RNA metabolism during infection. Dysregulation of RNA-binding proteins and mRNA processing pathways is broadly relevant to cancer biology and neurological and immunological phenotypes, making ZC3H7B a useful target for mechanistic studies of gene expression control.

RoXaN CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZC3H7B expression without altering the underlying DNA sequence.

RoXaN CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZC3H7B 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 ZC3H7B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RoXaN expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZC3H7B locus and enabling the study of RoXaN-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RoXaN pathway restoration in tumor cells with silenced or reduced ZC3H7B expression.

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