RPRD1B CRISPR Activation Plasmid (h): sc-412911-ACT

RPRD1B encodes a CTD-interacting protein that associates with RNA polymerase II and contributes to coordination of transcription elongation and co‑transcriptional RNA processing. By engaging the phosphorylated C‑terminal domain of POLR2A, RPRD1B helps regulate transcriptional timing, promoter-proximal pausing, and mRNA maturation programs that influence cell-cycle control and stress-responsive gene expression. Altered expression or regulatory disruption of transcriptional modulators such as RPRD1B is relevant to disease biology where global transcriptional balance and genome maintenance pathways are perturbed, including cancer-associated transcriptional reprogramming. The gene is therefore useful for mechanistic studies linking Pol II CTD signaling to downstream transcriptome changes.

RPRD1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPRD1B expression without altering the underlying DNA sequence.

RPRD1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPRD1B 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 RPRD1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RPRD1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPRD1B locus and enabling the study of RPRD1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RPRD1B pathway restoration in tumor cells with silenced or reduced RPRD1B expression.

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