DCST2 CRISPR Activation Plasmid (h): sc-405719-ACT

DCST2 (DC-STAMP domain-containing protein 2) is a multi-pass membrane protein implicated in cell–cell interaction processes and membrane fusion-related events, with strongest functional evidence coming from roles in gamete interaction and fertilization biology. In human tissues, DCST2 expression patterns support investigation into reproductive tract and germ cell regulatory programs, including membrane organization and signaling at specialized cell junctions. Although mechanistic details remain under active study, altered expression or regulation of DCST2 may be relevant to infertility-associated phenotypes and broader epithelial differentiation contexts. As a membrane-associated factor, DCST2 can be used to probe pathways linking surface protein composition to cellular communication and developmental biology.

DCST2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DCST2 expression without altering the underlying DNA sequence.

DCST2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DCST2 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 DCST2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DCST2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DCST2 locus and enabling the study of DCST2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DCST2 pathway restoration in tumor cells with silenced or reduced DCST2 expression.

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