UFD1 CRISPR Activation Plasmid (h): sc-403304-ACT

UFD1L encodes UFD1, a core component of the UFD1–NPL4–VCP/p97 complex that couples ubiquitin recognition to ATP-dependent extraction of proteins from membranes and macromolecular assemblies for proteasomal degradation. This axis supports endoplasmic reticulum–associated degradation (ERAD), ribosome-associated quality control, and resolution of stalled protein complexes, thereby maintaining proteostasis under stress. Through these processes, UFD1 influences cell cycle progression, genome stability, and responses to unfolded protein stress. Dysregulation of UFD1L-linked quality control pathways has been associated with altered stress tolerance and phenotypes observed in disorders involving proteostasis imbalance and chromosomal abnormalities, supporting its utility in mechanistic disease modeling.

UFD1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UFD1L expression without altering the underlying DNA sequence.

UFD1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UFD1L 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 UFD1L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UFD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UFD1L locus and enabling the study of UFD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UFD1 pathway restoration in tumor cells with silenced or reduced UFD1L expression.

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