Flotillin-2 CRISPR Activation Plasmid (h): sc-401200-ACT

Human FLOT2 encodes flotillin-2, a membrane-associated scaffolding protein enriched in lipid rafts and caveolae-like microdomains that organizes receptor complexes and influences membrane curvature and endocytic trafficking. Flotillin-2 participates in signal transduction linked to receptor tyrosine kinases and downstream MAPK/ERK and PI3K/AKT pathway activity, supporting regulation of cell adhesion, migration, and cytoskeletal dynamics. Through its role in membrane microdomain organization and vesicular transport, FLOT2 impacts exosome biology and spatial control of signaling nodes at the plasma membrane. Altered flotillin-2 expression or localization has been reported across multiple tumor types and other pathophysiology contexts where dysregulated signaling and membrane trafficking contribute to disease-associated phenotypes.

Flotillin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FLOT2 expression without altering the underlying DNA sequence.

Flotillin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FLOT2 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 FLOT2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Flotillin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FLOT2 locus and enabling the study of Flotillin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Flotillin-2 pathway restoration in tumor cells with silenced or reduced FLOT2 expression.

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