Psf2 CRISPR Activation Plasmid (h): sc-405023-ACT

Human GINS2 encodes Psf2, an essential subunit of the GINS complex that cooperates with CDC45 and the MCM2–7 helicase to form the CMG replisome required for DNA replication initiation and elongation. Psf2 supports origin firing, replication fork progression, and S-phase checkpoint integrity, linking its activity to genome stability maintenance and coordinated cell-cycle progression. Dysregulated GINS2 expression has been associated with proliferative programs and replication stress phenotypes observed across multiple disease research contexts, including cancer biology. As a core replication factor, Psf2 is frequently studied in pathways governing DNA damage responses, chromatin dynamics during S phase, and replication-associated transcriptional stress.

Psf2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GINS2 expression without altering the underlying DNA sequence.

Psf2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GINS2 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 GINS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Psf2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GINS2 locus and enabling the study of Psf2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Psf2 pathway restoration in tumor cells with silenced or reduced GINS2 expression.

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