SSRP1 CRISPR Activation Plasmid (h): sc-401780-ACT

SSRP1 (structure specific recognition protein 1) is a core subunit of the FACT (facilitates chromatin transcription) complex that modulates nucleosome disassembly and reassembly during transcription elongation, DNA replication, and DNA repair. By coordinating histone chaperone activity with RNA polymerase II progression, SSRP1 helps maintain chromatin accessibility and genome stability, linking it to pathways that govern replication stress responses and DNA damage signaling. Altered SSRP1 expression or FACT function has been associated with dysregulated transcriptional programs, proliferation, and chromatin architecture changes observed across multiple tumor contexts. As a chromatin regulator, SSRP1 is frequently studied in epigenetic control of gene expression, cell-cycle progression, and mechanisms of sensitivity to genotoxic stress in human cell models.

SSRP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SSRP1 expression without altering the underlying DNA sequence.

SSRP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SSRP1 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 SSRP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SSRP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SSRP1 locus and enabling the study of SSRP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SSRP1 pathway restoration in tumor cells with silenced or reduced SSRP1 expression.

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