CCNB1/cyclin B1 CRISPR Activation Plasmid (h): sc-400114-ACT

CCNB1 encodes cyclin B1, a core regulatory subunit of the CDK1 complex that drives the G2/M transition and coordinates entry into mitosis. Cyclin B1 accumulation and nuclear translocation promote chromosome condensation, nuclear envelope breakdown, and mitotic spindle assembly, while timely ubiquitin-mediated degradation by the APC/C supports mitotic exit. CCNB1 activity integrates with DNA damage checkpoints and cell-cycle surveillance pathways to ensure faithful genome replication and segregation. Dysregulated CCNB1 expression is frequently associated with aberrant proliferation, chromosomal instability, and tumor-associated cell-cycle phenotypes, making it a widely used marker and mechanistic node in mitotic control studies.

CCNB1/cyclin B1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCNB1 expression without altering the underlying DNA sequence.

CCNB1/cyclin B1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCNB1 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 CCNB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CCNB1/cyclin B1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCNB1 locus and enabling the study of CCNB1/cyclin B1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CCNB1/cyclin B1 pathway restoration in tumor cells with silenced or reduced CCNB1 expression.

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