UFM1 CRISPR Activation Plasmid (h): sc-416900-ACT

Human UFM1 encodes ubiquitin-fold modifier 1, a ubiquitin-like protein that is conjugated to substrates through the UFMylation cascade involving UBA5, UFC1, and UFL1. This post-translational modification regulates endoplasmic reticulum proteostasis, ribosome-associated quality control, and cellular responses to proteotoxic and oxidative stress, with established connections to ER homeostasis and secretory pathway function. UFM1-dependent signaling has been implicated in neurodevelopmental and hematologic phenotypes, and altered UFMylation activity is studied in the context of cancer cell survival and stress adaptation. As a core modifier in this pathway, UFM1 serves as a useful node for dissecting how ubiquitin-like conjugation tunes protein turnover, translation control, and organelle stress responses.

UFM1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UFM1 expression without altering the underlying DNA sequence.

UFM1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UFM1 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 UFM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UFM1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UFM1 locus and enabling the study of UFM1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UFM1 pathway restoration in tumor cells with silenced or reduced UFM1 expression.

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