CPSF4 CRISPR Activation Plasmid (h): sc-403190-ACT

Human CPSF4 (cleavage and polyadenylation specificity factor subunit 4) is an essential component of the CPSF complex that recognizes the AAUAAA polyadenylation signal and coordinates 3′ end cleavage and poly(A) tail addition on pre-mRNAs. Through coupling transcription termination with mRNA maturation, CPSF4 helps shape transcript stability, nuclear export, and translation efficiency, linking RNA processing to global gene expression control. Perturbation of CPSF4-dependent polyadenylation can shift alternative poly(A) site usage and alter isoform balance, with downstream effects on cell-cycle programs, stress responses, and differentiation. Dysregulated 3′ end processing and polyadenylation landscapes are recurrent features of cancer and other proliferative or RNA-metabolism–associated disorders, making CPSF4 a useful node for mechanistic studies of RNA processing–driven phenotypes.

CPSF4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CPSF4 expression without altering the underlying DNA sequence.

CPSF4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CPSF4 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 CPSF4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CPSF4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CPSF4 locus and enabling the study of CPSF4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CPSF4 pathway restoration in tumor cells with silenced or reduced CPSF4 expression.

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