CDKN1B/Kip1 p27 CRISPR Activation Plasmid (h): sc-400074-ACT

CDKN1B encodes p27^Kip1, a cyclin-dependent kinase inhibitor that restrains CDK2- and CDK4/6-driven progression through the G1/S checkpoint by modulating cyclin E/A–CDK2 and cyclin D–CDK4/6 complexes. p27 integrates mitogenic and antiproliferative cues downstream of pathways such as PI3K–AKT and TGF-β, and its abundance is tightly controlled by phosphorylation-dependent ubiquitination and proteasomal turnover. Through these mechanisms, CDKN1B influences proliferation, quiescence, and differentiation programs across diverse cell types. Altered CDKN1B expression or mislocalization is frequently linked to dysregulated cell-cycle control in tumor biology and to perturbations in endocrine and developmental contexts.

CDKN1B/Kip1 p27 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDKN1B expression without altering the underlying DNA sequence.

CDKN1B/Kip1 p27 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDKN1B 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 CDKN1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CDKN1B/Kip1 p27 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDKN1B locus and enabling the study of CDKN1B/Kip1 p27-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CDKN1B/Kip1 p27 pathway restoration in tumor cells with silenced or reduced CDKN1B expression.

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