N-WASP CRISPR Activation Plasmid (h): sc-401440-ACT

WASL encodes human N-WASP, a multidomain actin regulatory protein that couples Rho-family GTPase and phosphoinositide signaling to Arp2/3-mediated branched actin nucleation. By coordinating cytoskeletal remodeling, N-WASP supports processes including clathrin-mediated endocytosis, membrane protrusion and migration, adhesion dynamics, and immunological synapse organization. N-WASP activity integrates signaling inputs from Cdc42, SH3-domain adaptors, and tyrosine kinases to tune actin assembly in response to extracellular cues. Dysregulated WASL/N-WASP function has been implicated in altered cell motility and invasive behavior, neurodevelopmental phenotypes linked to synaptic actin dynamics, and immune cell signaling defects relevant to inflammation and host–pathogen interactions.

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

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

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