WRN CRISPR Activation Plasmid (h): sc-401860-ACT

Human WRN encodes the Werner syndrome ATP-dependent helicase, a RecQ family enzyme with 3′–5′ DNA helicase and exonuclease activities that safeguard genome stability. WRN participates in DNA replication, recombination, telomere maintenance, and multiple DNA damage response pathways, including processing of stalled replication forks and repair of double-strand breaks. Loss-of-function variants disrupt replication-associated repair and accelerate genomic instability, linking WRN dysfunction to Werner syndrome and broader mechanisms of premature aging and cancer predisposition. WRN is also a key node for studying replication stress signaling, telomere dynamics, and pathway dependencies in DNA repair–proficient versus –deficient backgrounds.

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

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

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