ICK CRISPR Activation Plasmid (h): sc-406371-ACT

Intestinal cell kinase (ICK) is a serine/threonine protein kinase that localizes to primary cilia and the nucleus, where it regulates cell-cycle progression, ciliogenesis, and microtubule-dependent processes. In human cells, ICK contributes to signaling networks linked to ciliary function and developmental patterning, including pathways that intersect with Hedgehog signal transduction and centrosome dynamics. Perturbation of ICK activity or expression is associated with ciliopathy-related phenotypes and developmental disorders, consistent with its role in coordinating proliferation with cilia-mediated signaling. ICK is also studied in contexts where altered kinase signaling and ciliary control influence differentiation states and cellular fitness.

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

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

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