TSPAN1 CRISPR Activation Plasmid (h): sc-404815-ACT

TSPAN1 (tetraspanin 1) is a four-pass transmembrane protein that organizes tetraspanin-enriched microdomains and modulates the spatial distribution of partner receptors, integrins, and signaling complexes at the cell surface. Through these assemblies, TSPAN1 can influence cell adhesion, motility, membrane trafficking, and proliferative signaling, linking it to processes such as epithelial organization and dynamic remodeling of the plasma membrane. Dysregulated TSPAN1 expression has been reported across multiple tumor contexts and is frequently studied in relation to invasion, metastasis-associated phenotypes, and changes in growth-factor pathway responsiveness. Its membrane localization and scaffolding role make it a useful node for investigating receptor cross-talk and downstream pathway rewiring in human cell models.

TSPAN1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TSPAN1 expression without altering the underlying DNA sequence.

TSPAN1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TSPAN1 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 TSPAN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TSPAN1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TSPAN1 locus and enabling the study of TSPAN1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TSPAN1 pathway restoration in tumor cells with silenced or reduced TSPAN1 expression.

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