cyclin D1 CRISPR Activation Plasmid (h): sc-400047-ACT

CCND1 encodes cyclin D1, a core regulator of the G1/S cell-cycle transition that binds and activates CDK4/6 to promote phosphorylation of RB family proteins and release of E2F-driven transcription. Through integration of mitogenic signaling from pathways such as MAPK/ERK and PI3K/AKT, cyclin D1 coordinates proliferative cues with cell-cycle entry and contributes to control of DNA replication and checkpoint responses. Dysregulated CCND1 expression or amplification is frequently associated with aberrant proliferation and genomic instability in diverse cancers, and altered cyclin D1 activity has also been linked to changes in differentiation programs and cellular metabolism. As a transcriptionally responsive cell-cycle node, CCND1 is widely used to interrogate signaling-to-proliferation coupling and oncogenic pathway dependencies in human cell models.

cyclin D1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCND1 expression without altering the underlying DNA sequence.

cyclin D1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCND1 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 CCND1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous cyclin D1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCND1 locus and enabling the study of cyclin D1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of cyclin D1 pathway restoration in tumor cells with silenced or reduced CCND1 expression.

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