NSFL1C p47 CRISPR Activation Plasmid (h): sc-402328-ACT

NSFL1C (p47) encodes an adaptor protein that binds the AAA+ ATPase VCP/p97 and helps target it to specific substrates in ubiquitin-dependent protein quality control. NSFL1C p47 is implicated in membrane trafficking and Golgi/ER dynamics, including p97-cofactor–mediated remodeling events that influence vesicle fusion and organelle reassembly. Through its role in proteostasis and stress-responsive degradation pathways such as ER-associated degradation, altered p47 function can modulate cellular sensitivity to proteotoxic stress and remodeling of secretory compartments. Dysregulation of the p97–cofactor axis has been linked to neurodegeneration and cancer-associated proteostasis dependencies, making NSFL1C a useful node for mechanistic studies of these processes.

NSFL1C p47 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NSFL1C expression without altering the underlying DNA sequence.

NSFL1C p47 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NSFL1C 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 NSFL1C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NSFL1C p47 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NSFL1C locus and enabling the study of NSFL1C p47-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NSFL1C p47 pathway restoration in tumor cells with silenced or reduced NSFL1C expression.

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