VPS13A CRISPR Activation Plasmid (h): sc-407168-ACT

VPS13A encodes vacuolar protein sorting-associated protein 13A, a large lipid-transfer factor that localizes at membrane contact sites and supports lipid homeostasis, organelle dynamics, and vesicular trafficking. VPS13A is implicated in mitochondria–endoplasmic reticulum communication, autophagy-related processes, and maintenance of neuronal and erythrocyte membrane integrity through regulation of phospholipid distribution. Disruption of VPS13A function is linked to neurodegenerative phenotypes and movement-disorder biology, including the pathogenesis of chorea-acanthocytosis, and it is increasingly studied in contexts of cellular stress responses and membrane remodeling. As a result, VPS13A provides a mechanistic entry point for interrogating pathways that couple intracellular transport to mitochondrial function and neurobiology.

VPS13A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VPS13A expression without altering the underlying DNA sequence.

VPS13A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VPS13A 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 VPS13A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous VPS13A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VPS13A locus and enabling the study of VPS13A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of VPS13A pathway restoration in tumor cells with silenced or reduced VPS13A expression.

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