WDR1 CRISPR Activation Plasmid (h): sc-402871-ACT

WDR1 (WD repeat domain 1), also known as AIP1, is an actin-regulatory protein that cooperates with cofilin to promote actin filament disassembly and turnover, supporting dynamic cytoskeletal remodeling. By controlling F-actin organization, WDR1 contributes to processes including cell migration, endocytosis, and immune-cell activation where rapid actin reorganization is required. Altered WDR1 function has been linked to dysregulated inflammatory signaling and immune phenotypes, consistent with its role in maintaining actin homeostasis in hematopoietic and other cell types. In human cell models, WDR1 is frequently studied to connect cytoskeletal dynamics with stress responses and signaling outputs that depend on actin-dependent trafficking and membrane remodeling.

WDR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous WDR1 expression without altering the underlying DNA sequence.

WDR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WDR1 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 WDR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous WDR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WDR1 locus and enabling the study of WDR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of WDR1 pathway restoration in tumor cells with silenced or reduced WDR1 expression.

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