MOK CRISPR Activation Plasmid (h): sc-410973-ACT

Human MOK (MAPK/MAK/MRK overlapping kinase) encodes a serine/threonine protein kinase within the MAPK-related CMGC kinase family that contributes to regulation of cellular signaling and differentiation programs. MOK activity has been linked to control of gene expression and cytoskeletal or junctional organization in specialized epithelia, supporting roles in tissue homeostasis and developmental processes. As a kinase node that can intersect with MAPK-associated networks, altered MOK expression or signaling is used in research to interrogate pathway rewiring that accompanies changes in proliferation, polarity, and stress responses. Dysregulated kinase expression patterns, including MOK, are studied for associations with oncogenic signaling contexts and other disorders where epithelial differentiation and signaling balance are perturbed.

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

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

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