ODR4 CRISPR Activation Plasmid (h): sc-417633-ACT

Human C1orf27 encodes ODR4, a conserved multi-pass membrane protein implicated in endoplasmic reticulum and secretory pathway quality control, where it supports the maturation and trafficking of select G protein–coupled receptors and other client proteins. By influencing receptor biogenesis and membrane targeting, ODR4 can modulate downstream signal transduction networks coupled to GPCR activity, including pathways affecting cellular responsiveness to extracellular cues. Altered proteostasis and receptor trafficking are broadly relevant to stress signaling, differentiation, and neuronal and endocrine functions, making ODR4 a useful node for dissecting membrane protein handling. Dysregulation of these processes is frequently observed across complex disease contexts, supporting investigation of ODR4 in models of signaling imbalance and cellular stress without implying clinical outcomes.

ODR4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous C1orf27 expression without altering the underlying DNA sequence.

ODR4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C1orf27 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 C1orf27 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ODR4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C1orf27 locus and enabling the study of ODR4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ODR4 pathway restoration in tumor cells with silenced or reduced C1orf27 expression.

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