Sel-1L CRISPR Activation Plasmid (h): sc-403861-ACT

SEL1L encodes Sel-1L, a core component of the endoplasmic reticulum–associated degradation (ERAD) machinery that partners with HRD1 to recognize, retrotranslocate, and promote ubiquitin-dependent proteasomal clearance of misfolded secretory and membrane proteins. Through this role it supports ER proteostasis, modulates unfolded protein response signaling, and influences the stability of diverse substrates involved in differentiation and stress adaptation. SEL1L activity intersects with pathways controlling ER quality control, ubiquitin ligase complexes, and secretory pathway homeostasis, processes frequently perturbed in conditions characterized by chronic ER stress. Dysregulated ERAD capacity and altered SEL1L expression have been explored in studies of neurodegeneration, metabolic dysfunction, and tumor biology as mechanisms linking proteotoxic stress to cell fate decisions.

Sel-1L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SEL1L expression without altering the underlying DNA sequence.

Sel-1L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SEL1L 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 SEL1L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sel-1L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SEL1L locus and enabling the study of Sel-1L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sel-1L pathway restoration in tumor cells with silenced or reduced SEL1L expression.

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