LASS4 CRISPR Activation Plasmid (h): sc-406329-ACT

CERS4 (LASS4) encodes ceramide synthase 4, an endoplasmic reticulum membrane enzyme that acylates sphingoid bases to generate specific long-chain ceramide species. By controlling ceramide composition, LASS4 influences sphingolipid-dependent membrane organization and signaling processes linked to apoptosis, cellular stress responses, and metabolic homeostasis. Altered ceramide synthesis and sphingolipid remodeling have been associated with dysregulated proliferation, inflammation, and neuro-metabolic phenotypes, making CERS4 a useful node for studying lipid signaling and organelle stress pathways. Functional interrogation of CERS4 supports mechanistic studies of ceramide-driven regulation of cell fate, barrier function, and lipid-mediated signal transduction.

LASS4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CERS4 expression without altering the underlying DNA sequence.

LASS4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CERS4 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 CERS4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LASS4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CERS4 locus and enabling the study of LASS4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LASS4 pathway restoration in tumor cells with silenced or reduced CERS4 expression.

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