Mnk1 CRISPR Activation Plasmid (h): sc-402417-ACT

Human MKNK1 encodes MAP kinase–interacting serine/threonine-protein kinase 1 (Mnk1), a downstream effector of ERK and p38 MAPK signaling that links extracellular cues to translational control. Mnk1 phosphorylates eIF4E and modulates cap-dependent mRNA translation, influencing the expression of growth- and stress-responsive transcripts involved in proliferation, survival, and inflammatory programs. Through these pathways, MKNK1 is frequently studied in contexts where MAPK-driven remodeling of the translatome contributes to altered cell-state decisions. Dysregulated MKNK1–eIF4E signaling has been associated with oncogenic signaling networks and immune/inflammatory regulation, making it a useful node for mechanistic studies of signaling-to-translation coupling.

Mnk1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MKNK1 expression without altering the underlying DNA sequence.

Mnk1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MKNK1 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 MKNK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mnk1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MKNK1 locus and enabling the study of Mnk1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mnk1 pathway restoration in tumor cells with silenced or reduced MKNK1 expression.

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