PSF CRISPR Activation Plasmid (h): sc-402024-ACT

Human SFPQ encodes the PSF protein, a multifunctional nuclear RNA- and DNA-binding factor that coordinates transcriptional regulation, pre-mRNA splicing, and RNA 3′-end processing. PSF participates in paraspeckle and nuclear body dynamics through interactions with long noncoding RNAs and RNA-binding partners, shaping gene expression programs under stress and differentiation cues. It also contributes to genome maintenance by influencing DNA damage response signaling and repair pathway choice, linking RNA processing to chromatin-associated transactions. Dysregulation of SFPQ/PSF-associated ribonucleoprotein complexes has been reported in contexts involving altered splicing fidelity, genomic instability, and aberrant transcriptional control that are frequently explored in cancer and neurodegeneration research.

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

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

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