CD9 CRISPR Activation Plasmid (h): sc-400252-ACT

Human CD9 encodes a tetraspanin cell-surface protein that organizes tetraspanin-enriched microdomains and modulates the distribution and signaling of partner receptors, including integrins and growth factor receptors. Through these membrane assemblies, CD9 influences cell adhesion, migration, membrane fusion events, and extracellular vesicle biogenesis, shaping pathways linked to cytoskeletal dynamics and intercellular communication. CD9 is widely used as a marker associated with exosomes and has been implicated in processes relevant to inflammation, fibrosis, and tumor cell dissemination via effects on cell–cell and cell–matrix interactions. Perturbation of CD9 expression is therefore informative for dissecting how membrane scaffolding impacts signaling specificity and cellular phenotypes in human model systems.

CD9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD9 expression without altering the underlying DNA sequence.

CD9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD9 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 CD9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD9 locus and enabling the study of CD9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD9 pathway restoration in tumor cells with silenced or reduced CD9 expression.

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