ARV1 CRISPR Activation Plasmid (h): sc-407460-ACT

ARV1 (ARV1 homolog) encodes an evolutionarily conserved, multi-pass membrane protein predominantly localized to the endoplasmic reticulum and implicated in lipid homeostasis. Human ARV1 has been linked to sterol and sphingolipid trafficking, maintenance of membrane composition, and coordination of ER-dependent processes that influence secretory pathway function and cellular stress responses. Through its role in organizing lipid balance and membrane dynamics, ARV1 can impact signaling networks coupled to organelle integrity and proteostasis. Perturbation of ARV1 expression has been associated with neurodevelopmental and epileptic phenotypes in genetic studies, supporting its relevance for disease-mechanism research in metabolic and neurological contexts.

ARV1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARV1 expression without altering the underlying DNA sequence.

ARV1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARV1 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 ARV1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARV1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARV1 locus and enabling the study of ARV1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARV1 pathway restoration in tumor cells with silenced or reduced ARV1 expression.

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