SPTLC1 CRISPR Activation Plasmid (h): sc-402829-ACT

SPTLC1 encodes a catalytic subunit of serine palmitoyltransferase, the rate-limiting enzyme for de novo sphingolipid biosynthesis that condenses serine with palmitoyl-CoA to generate 3-ketosphinganine. Through control of ceramide, sphingomyelin, and complex glycosphingolipid production, SPTLC1 influences membrane composition, lipid raft signaling, autophagy, and stress-response pathways including ER homeostasis. Altered SPTLC1 activity perturbs sphingolipid profiles and has been associated with neurodegenerative and peripheral neuropathy phenotypes as well as broader metabolic dysregulation. As a central node in lipid metabolism, SPTLC1 is widely studied for its impact on cellular signaling, organelle function, and lipid-driven inflammatory processes.

SPTLC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPTLC1 expression without altering the underlying DNA sequence.

SPTLC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPTLC1 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 SPTLC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPTLC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPTLC1 locus and enabling the study of SPTLC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPTLC1 pathway restoration in tumor cells with silenced or reduced SPTLC1 expression.

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