WAPL CRISPR Activation Plasmid (m): sc-432331-ACT

Mouse WAPL (Wapl) encodes a cohesin release factor that regulates the dynamic association of cohesin with chromatin, shaping higher-order genome organization and sister chromatid cohesion. By promoting cohesin unloading during interphase and mitotic progression, WAPL influences DNA replication timing, transcriptional regulation, and accurate chromosome segregation. WAPL activity intersects with DNA damage response and repair processes by modulating cohesin-dependent stabilization of stalled replication forks and repair foci. Dysregulated cohesin dynamics and altered WAPL expression have been associated with genome instability and cohesinopathy-linked phenotypes, making Wapl a useful node for studying chromatin architecture and cell-cycle control in disease-relevant models.

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

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

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