SMS1 CRISPR Activation Plasmid (h): sc-403382-ACT

Human SGMS1 encodes sphingomyelin synthase 1 (SMS1), a Golgi-localized enzyme that converts ceramide and phosphatidylcholine into sphingomyelin and diacylglycerol, thereby linking sphingolipid metabolism with lipid signaling. By regulating the balance between ceramide, sphingomyelin, and DAG, SMS1 influences membrane composition, lipid raft organization, vesicular trafficking, and downstream pathways affecting cell stress responses and proliferation. Perturbations in sphingomyelin homeostasis are associated with altered inflammatory signaling, metabolic dysfunction, and oncogenic phenotypes in model systems, making SGMS1 a useful node for dissecting lipid-driven cell biology. SMS1 activity is also relevant to studies of membrane biophysics and organelle communication due to its impact on Golgi and plasma membrane lipid remodeling.

SMS1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SGMS1 expression without altering the underlying DNA sequence.

SMS1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SGMS1 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 SGMS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMS1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SGMS1 locus and enabling the study of SMS1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMS1 pathway restoration in tumor cells with silenced or reduced SGMS1 expression.

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