Nek1 CRISPR Activation Plasmid (h): sc-403501-ACT

NEK1 encodes NIMA-related kinase 1 (Nek1), a serine/threonine kinase implicated in cell-cycle control, centrosome and cilia biology, and coordination of the DNA damage response. Nek1 has been linked to signaling processes that regulate microtubule dynamics, checkpoint signaling, and genome stability through phosphorylation-dependent control of key mitotic and repair-associated substrates. In human cells, altered NEK1 activity is associated with defects in ciliogenesis and impaired responses to genotoxic stress, connecting this kinase to pathways that shape cellular homeostasis. Genetic and functional studies have also implicated NEK1 in disease-relevant mechanisms, including neurodegeneration and cancer-related genome instability phenotypes.

Nek1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NEK1 expression without altering the underlying DNA sequence.

Nek1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NEK1 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 NEK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nek1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NEK1 locus and enabling the study of Nek1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nek1 pathway restoration in tumor cells with silenced or reduced NEK1 expression.

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