SPNS2 CRISPR Activation Plasmid (m): sc-432145-ACT

Mouse Spns2 encodes SPNS2, a multipass membrane transporter that exports sphingosine-1-phosphate (S1P) to establish extracellular S1P gradients that shape S1P receptor signaling. By controlling S1P availability, SPNS2 influences lymphocyte trafficking, endothelial barrier regulation, angiogenic cues, and inflammatory cell recruitment within the broader sphingolipid metabolic network. Perturbation of SPNS2-mediated S1P transport has been linked to altered immune homeostasis and vascular biology, making Spns2 a useful node for studying immunoregulation, inflammation-associated phenotypes, and microenvironmental signaling. In mouse models, Spns2 activity is commonly investigated in the context of tissue-specific S1P flux, immune cell egress, and vascular development pathways.

SPNS2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Spns2 expression without altering the underlying DNA sequence.

SPNS2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Spns2 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 Spns2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPNS2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Spns2 locus and enabling the study of SPNS2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPNS2 pathway restoration in tumor cells with silenced or reduced Spns2 expression.

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