UFL1 CRISPR Activation Plasmid (h): sc-407950-ACT

UFL1 encodes an E3 ligase that catalyzes UFMylation, a ubiquitin-like modification pathway important for endoplasmic reticulum homeostasis and proteostasis. By coordinating conjugation of UFM1 to specific substrates, UFL1 influences ER stress signaling, protein quality control, and pathways linked to ribosome-associated surveillance and secretory protein handling. Altered UFL1 activity has been associated in the literature with dysregulated cellular stress responses and inflammatory signaling, providing a mechanistic connection to disease-relevant phenotypes in tissues with high protein-folding demand. As a result, UFL1 is frequently studied in models probing ER stress adaptation, cell survival under proteotoxic stress, and pathway crosstalk between ubiquitin-like modifiers and innate immune regulation.

UFL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UFL1 expression without altering the underlying DNA sequence.

UFL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UFL1 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 UFL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UFL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UFL1 locus and enabling the study of UFL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UFL1 pathway restoration in tumor cells with silenced or reduced UFL1 expression.

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