MCM5 CRISPR Activation Plasmid (h): sc-401980-ACT

MCM5 encodes a core subunit of the minichromosome maintenance (MCM2–7) helicase complex that licenses replication origins and drives DNA unwinding during S phase. By coordinating origin firing and replication fork progression, MCM5 helps maintain genome stability and supports orderly cell-cycle transitions through DNA replication and checkpoint control pathways. Dysregulated MCM5 expression is frequently associated with hyperproliferative states and replication stress signatures observed in multiple cancer and genomic instability research contexts. Its central role in DNA synthesis also makes MCM5 a useful node for studying replication–transcription conflicts and cellular responses to stalled forks.

MCM5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MCM5 expression without altering the underlying DNA sequence.

MCM5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MCM5 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 MCM5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MCM5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MCM5 locus and enabling the study of MCM5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MCM5 pathway restoration in tumor cells with silenced or reduced MCM5 expression.

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