PRHOXNB CRISPR Activation Plasmid (h): sc-416145-ACT

Human URAD encodes the PRHOXNB protein, a putative transcriptional regulator implicated in controlling gene expression programs that shape cellular identity and state transitions. By influencing promoter and enhancer activity, PRHOXNB is expected to intersect with core transcriptional and chromatin-regulatory processes that coordinate proliferation, differentiation, and stress-adaptive responses. Aberrant regulation of transcription factor networks is a recurrent feature of complex disease biology, making URAD/PRHOXNB a relevant target for mechanistic studies of pathway rewiring and phenotype plasticity. Modulating URAD expression provides a route to interrogate downstream gene networks and their contributions to context-dependent cellular behaviors.

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

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

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