WASP CRISPR Activation Plasmid (h): sc-400712-ACT

Human WAS encodes Wiskott–Aldrich syndrome protein (WASP), a hematopoietic cell–restricted regulator of actin cytoskeleton remodeling that links receptor-proximal signaling to Arp2/3-dependent actin nucleation. Through interactions with Cdc42, phosphoinositides, and SH3-domain adaptors, WASP coordinates immune synapse formation, chemotaxis, phagocytosis, and vesicular trafficking, shaping T cell and B cell activation as well as innate immune effector functions. WASP activity integrates signaling from immunoreceptors and integrins to control cytoskeletal dynamics and transcriptional outputs during immune responses. Disrupted WAS function is associated with immunodeficiency and dysregulated inflammation, making WAS/WASP a key target for mechanistic studies of cytoskeleton-driven signaling in human immune cells.

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

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

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