SNX27 CRISPR Activation Plasmid (h): sc-403237-ACT

SNX27 (sorting nexin 27) is a PX domain–containing endosomal adaptor that couples membrane phosphoinositide recognition to cargo recycling, primarily through its PDZ domain interaction with PDZ-binding motifs on transmembrane proteins. By associating with the retromer and WASH complexes, SNX27 helps coordinate endosome-to-plasma membrane trafficking and actin-dependent tubulation, shaping receptor availability and downstream signaling. These processes influence synaptic receptor homeostasis, nutrient transporter recycling, and maintenance of epithelial polarity, linking SNX27 to pathways controlling cellular signaling amplitude and membrane proteostasis. Altered SNX27 expression or function has been implicated in neurodevelopmental and neurodegenerative phenotypes and in dysregulated receptor trafficking observed across several disease-relevant cellular contexts.

SNX27 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNX27 expression without altering the underlying DNA sequence.

SNX27 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNX27 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 SNX27 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SNX27 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNX27 locus and enabling the study of SNX27-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SNX27 pathway restoration in tumor cells with silenced or reduced SNX27 expression.

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