casein kinase Iδ CRISPR Activation Plasmid (h): sc-401839-ACT

CSNK1D encodes human casein kinase Iδ (CK1δ), a serine/threonine protein kinase that phosphorylates diverse substrates to coordinate circadian clock timing, cell-cycle progression, DNA damage responses, and vesicular trafficking. CK1δ contributes to signal transduction networks including Wnt/β-catenin and Hedgehog pathways through phosphorylation-dependent control of substrate stability, localization, and turnover. By modulating phosphorylation of clock and signaling proteins, CSNK1D influences transcriptional rhythms and proteostasis, processes frequently perturbed in proliferative and neurobiological disorders. Dysregulated CK1δ activity has been linked to altered circadian phenotypes and aberrant signaling outputs that intersect with oncogenic and neurodegeneration-associated pathways.

casein kinase Iδ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CSNK1D expression without altering the underlying DNA sequence.

casein kinase Iδ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CSNK1D 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 CSNK1D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous casein kinase Iδ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CSNK1D locus and enabling the study of casein kinase Iδ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of casein kinase Iδ pathway restoration in tumor cells with silenced or reduced CSNK1D expression.

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