EVA1B CRISPR Activation Plasmid (h): sc-412477-ACT

EVA1B (also known as FAM176B) encodes a transmembrane protein localized primarily to intracellular membranes, where it has been linked to regulation of vesicular trafficking, autophagy-associated processes, and cell survival signaling. Emerging studies suggest EVA1B influences stress-response pathways that shape cellular homeostasis, with context-dependent effects on proliferation and inflammatory programs. Altered EVA1B expression has been reported across multiple disease-relevant settings, including cancer-associated transcriptional changes and immune-related phenotypes, supporting its use as a mechanistic node in pathway interrogation. In human cell models, modulating EVA1B provides a route to dissect how membrane-associated signaling interfaces with organelle dynamics and transcriptional adaptation.

EVA1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EVA1B expression without altering the underlying DNA sequence.

EVA1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EVA1B 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 EVA1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EVA1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EVA1B locus and enabling the study of EVA1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EVA1B pathway restoration in tumor cells with silenced or reduced EVA1B expression.

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