PRC1 CRISPR Activation Plasmid (h): sc-401679-ACT

PRC1 (protein regulator of cytokinesis 1) encodes a microtubule-associated protein that organizes the central spindle and midbody during late mitosis, coordinating spindle midzone formation and completion of cytokinesis. Its activity is regulated across the cell cycle and intersects with mitotic kinase signaling, including pathways that control microtubule bundling, anaphase progression, and abscission. Altered PRC1 expression or regulation can perturb chromosomal segregation and cytokinetic fidelity, processes linked to genomic instability and proliferative phenotypes observed in multiple cancer contexts. PRC1 is therefore widely used as a marker and mechanistic node for studying mitotic control, cell division timing, and microtubule dynamics in human cells.

PRC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRC1 expression without altering the underlying DNA sequence.

PRC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRC1 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 PRC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRC1 locus and enabling the study of PRC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRC1 pathway restoration in tumor cells with silenced or reduced PRC1 expression.

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