ACBD3 CRISPR Activation Plasmid (h): sc-404320-ACT

Human ACBD3 (acyl-CoA binding domain containing 3), also known as GCP60, is a Golgi-associated scaffolding protein that coordinates membrane trafficking and organelle communication. It interacts with phosphatidylinositol 4-kinase signaling machinery and multiple viral and host factors to modulate Golgi structure, lipid homeostasis, and replication organelle formation. Through roles in vesicle tethering, protein sorting, and lipid transfer at membrane contact sites, ACBD3 influences secretory pathway dynamics and cellular stress responses. Dysregulated ACBD3-associated pathways have been studied in contexts including viral infection biology and cancer-associated changes in Golgi function and signaling.

ACBD3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ACBD3 expression without altering the underlying DNA sequence.

ACBD3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ACBD3 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 ACBD3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACBD3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ACBD3 locus and enabling the study of ACBD3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACBD3 pathway restoration in tumor cells with silenced or reduced ACBD3 expression.

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