Lamellipodin CRISPR Activation Plasmid (h): sc-403034-ACT

RAPH1 encodes lamellipodin, an Ena/VASP- and actin-associated adaptor that couples phosphoinositide signaling to cytoskeletal remodeling at lamellipodia and membrane ruffles. By interacting with PI(3,4)P2/PI(3,4,5)P3-enriched membranes and actin regulators, lamellipodin supports Rac-driven protrusion dynamics, cell spreading, and directional migration, linking to pathways that coordinate adhesion turnover and endocytosis. Altered RAPH1 expression or lamellipodin-dependent motility programs have been associated with invasive phenotypes and dysregulated tissue remodeling in disease-relevant contexts, making it a useful node for studying migration-centric signaling networks in human cells. These functions are frequently interrogated in models of cancer cell dissemination, neurodevelopmental processes, and immune cell trafficking without implying clinical outcomes.

Lamellipodin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAPH1 expression without altering the underlying DNA sequence.

Lamellipodin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAPH1 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 RAPH1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Lamellipodin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAPH1 locus and enabling the study of Lamellipodin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Lamellipodin pathway restoration in tumor cells with silenced or reduced RAPH1 expression.

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