p57 Kip2 CRISPR Activation Plasmid (h): sc-400444-ACT

CDKN1C encodes p57 Kip2, an imprinted CIP/KIP family cyclin-dependent kinase inhibitor that restrains CDK2 and CDK1 activity to enforce G1 arrest and coordinate differentiation programs. p57 Kip2 integrates signals from developmental and growth factor pathways, including TGF-β/SMAD and PI3K–AKT axis inputs, to modulate cell-cycle checkpoints, lineage commitment, and tissue growth control. Dysregulated CDKN1C expression and imprinting defects are linked to aberrant proliferation and developmental disorders, and altered p57 Kip2 levels are frequently studied in contexts such as cancer biology, stem cell quiescence, and placental development. As a nuclear regulator of proliferation, p57 Kip2 is also relevant to epigenetic regulation at the 11p15 imprinting cluster and to pathways governing senescence and cell fate transitions.

p57 Kip2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDKN1C expression without altering the underlying DNA sequence.

p57 Kip2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDKN1C 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 CDKN1C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p57 Kip2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDKN1C locus and enabling the study of p57 Kip2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p57 Kip2 pathway restoration in tumor cells with silenced or reduced CDKN1C expression.

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