MFSD2 CRISPR Activation Plasmid (h): sc-405672-ACT

MFSD2A encodes MFSD2, a sodium-dependent transporter of lysophosphatidylcholines that mediates uptake of docosahexaenoic acid (DHA) across endothelial barriers, with prominent roles at the blood–brain barrier. By regulating lipid influx and membrane phospholipid composition, MFSD2A supports neurodevelopment, neuronal maintenance, and barrier integrity while integrating into broader lipid transport and metabolic homeostasis pathways. MFSD2A also contributes to suppression of vesicular transcytosis in brain endothelium, linking its activity to endothelial trafficking control and tight barrier function. Altered MFSD2A expression or function has been associated with neurodevelopmental phenotypes and dysregulated barrier permeability, making it relevant for studies of CNS lipid metabolism and endothelial biology.

MFSD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MFSD2A expression without altering the underlying DNA sequence.

MFSD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MFSD2A 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 MFSD2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MFSD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MFSD2A locus and enabling the study of MFSD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MFSD2 pathway restoration in tumor cells with silenced or reduced MFSD2A expression.

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