Cks1 CRISPR Activation Plasmid (h): sc-403019-ACT

CKS1B encodes Cks1, a conserved cyclin-dependent kinase regulatory subunit that coordinates cell-cycle progression by modulating CDK activity and substrate selection. Cks1 is closely linked to SCF(SKP2)-mediated ubiquitin–proteasome control of cell-cycle inhibitors such as CDKN1B/p27, thereby influencing the G1/S transition, replication competence, and checkpoint fidelity. Through these interactions, CKS1B contributes to proliferative signaling networks that intersect with DNA damage responses and mitotic control. Dysregulated CKS1B expression has been associated with aberrant proliferation and genomic instability phenotypes that are frequently examined in oncology and hematologic malignancy research contexts.

Cks1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CKS1B expression without altering the underlying DNA sequence.

Cks1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CKS1B 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 CKS1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cks1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CKS1B locus and enabling the study of Cks1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cks1 pathway restoration in tumor cells with silenced or reduced CKS1B expression.

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