3632451O06Rik CRISPR Activation Plasmid (m): sc-426549-ACT

Mouse 3632451O06Rik encodes a largely uncharacterized protein with limited functional annotation, making it a useful target for mapping gene-regulatory networks and context-dependent cellular phenotypes. Emerging transcript-level evidence suggests 3632451O06Rik may participate in cell-state control processes such as proliferation, stress adaptation, and differentiation, where modest expression changes can alter pathway output. Because the gene is not well integrated into curated signaling maps, controlled perturbation is valuable for linking 3632451O06Rik expression to downstream transcriptional programs and protein interaction modules. Such studies can support disease-relevant biology by identifying associations with aberrant gene expression signatures observed in inflammation, neurobiology, and cancer-model contexts without implying clinical utility.

3632451O06Rik CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous 3632451O06Rik expression without altering the underlying DNA sequence.

3632451O06Rik CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the 3632451O06Rik 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 3632451O06Rik transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous 3632451O06Rik expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native 3632451O06Rik locus and enabling the study of 3632451O06Rik-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of 3632451O06Rik pathway restoration in tumor cells with silenced or reduced 3632451O06Rik expression.

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