Rad52 CRISPR Activation Plasmid (h): sc-401948-ACT

Human RAD52 encodes Rad52, a DNA repair factor that promotes homologous recombination and single-strand annealing to restore genomic integrity following double-strand breaks. Rad52 participates in replication-associated repair and recombination-mediated restart, coordinating with DNA end resection and RAD51-dependent processes within the broader DNA damage response. Its activity influences chromosomal stability, recombination outcomes, and cellular sensitivity to genotoxic stress. Altered RAD52 function or dependency has been studied in the context of genome instability phenotypes observed across multiple cancer-associated repair backgrounds and in models of impaired homologous recombination.

Rad52 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAD52 expression without altering the underlying DNA sequence.

Rad52 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAD52 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 RAD52 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rad52 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAD52 locus and enabling the study of Rad52-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rad52 pathway restoration in tumor cells with silenced or reduced RAD52 expression.

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