CPEB4 CRISPR Activation Plasmid (h): sc-413379-ACT

Human CPEB4 (cytoplasmic polyadenylation element binding protein 4) is an RNA-binding regulator that controls cytoplasmic polyadenylation, mRNA stability, and translation through recognition of CPE sequences in 3′ UTRs. By coordinating post-transcriptional gene expression programs, CPEB4 influences cell-cycle progression, neuronal plasticity, and stress-adaptive responses, and interfaces with signaling networks that remodel translational output during differentiation and metabolic challenge. Dysregulated CPEB4 expression or activity has been linked to altered proteostasis and aberrant gene-expression signatures in cancer biology, neurodevelopmental and neurodegenerative contexts, and inflammatory phenotypes. These properties make CPEB4 a useful node for dissecting RNA regulons, translational control mechanisms, and pathway-level rewiring under disease-relevant conditions.

CPEB4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CPEB4 expression without altering the underlying DNA sequence.

CPEB4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CPEB4 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 CPEB4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CPEB4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CPEB4 locus and enabling the study of CPEB4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CPEB4 pathway restoration in tumor cells with silenced or reduced CPEB4 expression.

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