YDJC CRISPR Activation Plasmid (h2): sc-414675-ACT-2

Human YDJC encodes a conserved putative deacetylase/epimerase family protein implicated in cellular metabolite processing and maintenance of proteostasis, with evidence supporting roles in redox balance and stress-adaptive responses. YDJC has been linked to modulation of protein quality control and mitochondrial-associated homeostatic pathways, making it relevant for studies of oxidative stress, inflammation-associated signaling, and metabolic remodeling. Altered YDJC expression has been reported across multiple disease contexts, including cancer and neurodegeneration-associated molecular signatures, suggesting utility as a mechanistic node for investigating dysregulated cellular resilience. Gene editing of YDJC enables functional interrogation of its contribution to metabolic and stress-response networks, supporting experiments in pathway mapping, genotype–phenotype analysis, and model generation for systems-level studies of cell state regulation.

YDJC CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous YDJC expression without altering the underlying DNA sequence.

YDJC CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the YDJC 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 YDJC transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous YDJC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native YDJC locus and enabling the study of YDJC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of YDJC pathway restoration in tumor cells with silenced or reduced YDJC expression.

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