SMPDL3B CRISPR Activation Plasmid (m): sc-430315-ACT

Mouse Smpdl3b encodes sphingomyelin phosphodiesterase acid-like 3B (SMPDL3B), a membrane-associated phosphodiesterase-like protein implicated in regulating sphingolipid and phospholipid metabolism at the cell surface and in endolysosomal compartments. SMPDL3B has been linked to modulation of innate immune signaling and membrane microdomain organization, influencing processes such as receptor trafficking, inflammatory pathway tuning, and cytoskeletal dynamics. In myeloid and tissue-resident immune cells, altered SMPDL3B activity can shift lipid-dependent signaling thresholds that shape responses to microbial cues and tissue stress. Dysregulation of lipid handling and immune homeostasis makes Smpdl3b a relevant target for mechanistic studies in inflammation-associated phenotypes and metabolic-immune crosstalk in mouse models.

SMPDL3B CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Smpdl3b expression without altering the underlying DNA sequence.

SMPDL3B CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Smpdl3b 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 Smpdl3b transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMPDL3B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Smpdl3b locus and enabling the study of SMPDL3B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMPDL3B pathway restoration in tumor cells with silenced or reduced Smpdl3b expression.

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