SP-lyase Lentiviral Activation Particles (m): sc-422908-LAC

Mouse Sgpl1 encodes sphingosine-1-phosphate lyase (SP-lyase), an endoplasmic reticulum–associated enzyme that irreversibly cleaves sphingosine-1-phosphate (S1P) to phosphoethanolamine and hexadecenal, thereby terminating S1P signaling and regulating intracellular sphingolipid homeostasis. By controlling the balance between S1P, sphingosine, and ceramide pools, SP-lyase influences pathways governing cell survival, stress responses, and lipid-mediated signaling, including processes linked to immune cell trafficking and vascular biology. Altered SGPL1 activity has been associated with disrupted sphingolipid metabolism and phenotypes relevant to inflammation, kidney and adrenal dysfunction, and neurodevelopmental abnormalities, making Sgpl1 a useful node for mechanistic studies in metabolic and signaling networks.

SP-lyase Lentiviral Activation Particles (m) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient Sgpl1 upregulation across a broader range of human cell types.

SP-lyase Lentiviral Activation Particles (m) deliver all functional components of the synergistic activation mediator (SAM) system via lentiviral transduction. The system comprises three particle preparations co-transduced into target cells: one encoding catalytically inactive dCas9 (D10A and N863A mutations) fused to the VP64 transactivation domain with a blasticidin resistance gene; one encoding the MS2-p65-HSF1 fusion protein with a hygromycin resistance gene; and one encoding a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers with a puromycin resistance gene. Following lentiviral transduction and genomic integration of the expression cassettes, the SAM components are stably expressed and assemble at the target locus within the proximal promoter region upstream of the Sgpl1 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous SP-lyase expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native Sgpl1 genomic locus and regulatory architecture.

The lentiviral format offers several practical advantages: stable genomic integration supports heritable activation across cell divisions; high-titer particle preparations eliminate the need for in-house viral production; and compatibility with primary, non-dividing, and transfection-resistant cell types expands experimental accessibility. Successful transduction can be confirmed and enriched through triple antibiotic selection using puromycin, hygromycin, and blasticidin.

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