SNX19 CRISPR Activation Plasmid (h): sc-406007-ACT

Human SNX19 (sorting nexin 19) encodes a PX domain–containing sorting nexin implicated in phosphoinositide-dependent membrane association and endosomal trafficking. As part of the broader sorting nexin network, SNX19 is linked to endosome-to-Golgi and endosome-to-plasma membrane transport processes that influence receptor recycling, membrane protein turnover, and spatial organization of signaling. Through these roles in vesicular transport and membrane dynamics, altered SNX19 activity can impact cellular homeostasis pathways that are frequently perturbed in complex disease contexts, including dysregulated signaling and metabolic phenotypes. SNX19 is therefore of interest for mechanistic studies connecting intracellular trafficking to pathway modulation and cell state changes.

SNX19 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNX19 expression without altering the underlying DNA sequence.

SNX19 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNX19 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 SNX19 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SNX19 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNX19 locus and enabling the study of SNX19-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SNX19 pathway restoration in tumor cells with silenced or reduced SNX19 expression.

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