Synoviolin CRISPR Activation Plasmid (h): sc-403476-ACT

Human SYVN1 encodes synoviolin, an endoplasmic reticulum (ER)–resident RING-type E3 ubiquitin ligase that promotes protein quality control through ER-associated degradation (ERAD). Synoviolin catalyzes ubiquitination of misfolded or regulatory substrates, supporting proteostasis, modulating unfolded protein response signaling, and influencing inflammatory and stress-adaptive pathways. By regulating stability of key signaling and folding intermediates, SYVN1 links ER stress to apoptosis resistance, cytokine-driven responses, and metabolic homeostasis. Dysregulated SYVN1 activity has been associated with chronic inflammatory phenotypes and aberrant cell survival programs, making it a useful node for mechanistic studies of ER stress–inflammation coupling.

Synoviolin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SYVN1 expression without altering the underlying DNA sequence.

Synoviolin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SYVN1 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 SYVN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Synoviolin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SYVN1 locus and enabling the study of Synoviolin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Synoviolin pathway restoration in tumor cells with silenced or reduced SYVN1 expression.

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