SNX17 CRISPR Activation Plasmid (h): sc-403452-ACT

SNX17 (sorting nexin 17) is a PX domain–containing endosomal adaptor that binds phosphoinositides and recognizes NPxY motifs to control endocytic recycling of diverse membrane proteins. Through interactions with Retriever/CCC complexes and retromer-associated trafficking machinery, SNX17 supports receptor sorting from early endosomes back to the plasma membrane, thereby shaping signaling output, nutrient uptake, and cell–matrix communication. By regulating surface availability of cargo such as LDL receptor family members and integrins, SNX17 influences pathways linked to lipid homeostasis, migration, and cytoskeletal organization. Dysregulated SNX17-dependent trafficking has been associated with altered receptor signaling and has been studied in contexts including metabolic phenotypes, neurobiology, and cancer-related cell behavior.

SNX17 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNX17 expression without altering the underlying DNA sequence.

SNX17 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNX17 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 SNX17 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SNX17 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNX17 locus and enabling the study of SNX17-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SNX17 pathway restoration in tumor cells with silenced or reduced SNX17 expression.

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