UVRAG CRISPR Activation Plasmid (h): sc-402956-ACT

Human UVRAG (UV radiation resistance associated gene) encodes a multifunctional regulator of autophagy and endolysosomal trafficking that supports cellular homeostasis under stress. UVRAG interacts with class III PI3K complexes to promote autophagosome maturation and autophagosome–lysosome fusion, and it also contributes to endocytic trafficking and vesicle dynamics through interactions with tethering and fusion machinery. Beyond degradative pathways, UVRAG has been linked to maintenance of genomic integrity and coordination of damage responses, connecting vesicular systems to cell survival programs. Dysregulation of UVRAG-associated processes is frequently studied in the context of tumor biology, neurodegeneration, and infection models where altered autophagy and membrane trafficking can reshape metabolism, inflammation, and proteostasis.

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

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

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