cyclin D3 CRISPR Activation Plasmid (h): sc-400634-ACT

CCND3 encodes cyclin D3, a regulatory cyclin that complexes with CDK4/6 to phosphorylate RB family proteins and promote G1/S cell-cycle progression. Through control of E2F-dependent transcription and integration of mitogenic cues, cyclin D3 helps coordinate proliferation with differentiation programs in hematopoietic and lymphoid lineages. Dysregulated CCND3 expression or activity has been associated with altered cell-cycle control and is implicated in oncogenic signaling contexts where cyclin D–CDK activity is elevated. Consequently, CCND3 is widely studied in pathways governing proliferation, checkpoint regulation, and lineage-specific growth control.

cyclin D3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCND3 expression without altering the underlying DNA sequence.

cyclin D3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCND3 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 CCND3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous cyclin D3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCND3 locus and enabling the study of cyclin D3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of cyclin D3 pathway restoration in tumor cells with silenced or reduced CCND3 expression.

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