CPSF1 CRISPR Activation Plasmid (h): sc-405273-ACT

Human CPSF1 (cleavage and polyadenylation specificity factor subunit 1) is a core component of the 3′-end processing machinery that recognizes polyadenylation signals and coordinates pre-mRNA cleavage and poly(A) tail addition. By coupling transcription termination with RNA maturation, CPSF1 helps regulate mRNA stability, nuclear export, and translation, thereby shaping global gene expression programs. Disruption of cleavage and polyadenylation control can alter alternative polyadenylation patterns and downstream regulatory networks, processes frequently implicated in proliferative signaling and stress-response pathways. As a central node in RNA processing, CPSF1 is widely studied in the context of transcriptional regulation, cell-cycle control, and disease-associated transcriptome remodeling.

CPSF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CPSF1 expression without altering the underlying DNA sequence.

CPSF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CPSF1 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 CPSF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CPSF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CPSF1 locus and enabling the study of CPSF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CPSF1 pathway restoration in tumor cells with silenced or reduced CPSF1 expression.

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