SPTLC3 CRISPR Activation Plasmid (h): sc-407807-ACT

SPTLC3 encodes a catalytic subunit of serine palmitoyltransferase, the rate-limiting enzyme that initiates de novo sphingolipid biosynthesis by condensing serine with palmitoyl-CoA. By regulating levels of sphingoid bases, ceramides, and downstream complex sphingolipids, SPTLC3 influences membrane composition, lipid raft organization, and signaling processes linked to apoptosis, inflammation, and metabolic adaptation. SPTLC3 activity interfaces with endoplasmic reticulum lipid homeostasis and broader pathways connecting ceramide metabolism to cellular stress responses. Altered sphingolipid profiles and dysregulated SPTLC3 expression have been associated with cardiometabolic traits and inflammatory phenotypes, making it a useful node for mechanistic studies of lipid-driven signaling.

SPTLC3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPTLC3 expression without altering the underlying DNA sequence.

SPTLC3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPTLC3 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 SPTLC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPTLC3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPTLC3 locus and enabling the study of SPTLC3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPTLC3 pathway restoration in tumor cells with silenced or reduced SPTLC3 expression.

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