PABP CRISPR Activation Plasmid (h): sc-400688-ACT

Human PABPC1 encodes poly(A)-binding protein (PABP), a central regulator of cytoplasmic mRNA metabolism that binds 3′ poly(A) tails to promote translation initiation and stabilize transcripts. Through interactions with translation initiation factors and components of the mRNA decay machinery, PABP coordinates mRNA circularization, translational efficiency, and deadenylation-dependent turnover. PABPC1 also contributes to stress granule and processing body dynamics, linking translational control to cellular stress responses. Altered PABP activity and PABPC1 expression have been associated with dysregulated gene expression programs observed in cancer and virus–host interactions, making it relevant to studies of oncogenic signaling and innate immune modulation.

PABP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PABPC1 expression without altering the underlying DNA sequence.

PABP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PABPC1 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 PABPC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PABP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PABPC1 locus and enabling the study of PABP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PABP pathway restoration in tumor cells with silenced or reduced PABPC1 expression.

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