LASS2 CRISPR Activation Plasmid (h): sc-403112-ACT

Human CERS2 (also known as LASS2) encodes ceramide synthase 2, a key enzyme in de novo sphingolipid biosynthesis that preferentially generates very-long-chain ceramides. By controlling ceramide chain-length composition, LASS2 influences membrane biophysics, lipid raft organization, and downstream signaling processes linked to apoptosis, autophagy, and stress responses. CERS2-dependent sphingolipid remodeling integrates with ER homeostasis and mitochondrial function and can modulate pathways affecting cell growth, differentiation, and inflammatory signaling. Dysregulated ceramide metabolism involving CERS2 has been associated with cancer biology, metabolic dysfunction, and neurodegenerative or liver-related disease mechanisms in experimental systems.

LASS2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CERS2 expression without altering the underlying DNA sequence.

LASS2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CERS2 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 CERS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LASS2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CERS2 locus and enabling the study of LASS2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LASS2 pathway restoration in tumor cells with silenced or reduced CERS2 expression.

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