P15RS CRISPR Activation Plasmid (h): sc-403743-ACT

RPRD1A encodes the human protein P15RS, a CTD-interacting factor that associates with RNA polymerase II and contributes to coupling transcription with co-transcriptional RNA processing. Through modulation of polymerase CTD phosphorylation dynamics and interactions with transcriptional regulators, P15RS influences gene expression programs linked to cell-cycle control, DNA damage signaling, and chromatin-associated processes. Dysregulation of transcriptional control and RNA processing is a recurrent feature of proliferative and stress-response phenotypes, making RPRD1A a useful target for mechanistic studies in cancer biology and related gene expression disorders. Researchers commonly leverage RPRD1A perturbation to map transcriptional networks, interrogate RNA polymerase II–dependent pathways, and profile downstream transcriptomic and proteomic changes.

P15RS CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPRD1A expression without altering the underlying DNA sequence.

P15RS CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPRD1A 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 RPRD1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous P15RS expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPRD1A locus and enabling the study of P15RS-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of P15RS pathway restoration in tumor cells with silenced or reduced RPRD1A expression.

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