MLK3 CRISPR Activation Plasmid (h): sc-401781-ACT

MAP3K11 encodes the human mixed lineage kinase 3 (MLK3), a MAP3K that couples diverse upstream stress and cytokine cues to downstream MAPK signaling, including the JNK and p38 cascades. Through phosphorylation of MAP2Ks such as MKK4/7 and MKK3/6, MLK3 helps regulate transcriptional programs controlling inflammation, apoptosis, cytoskeletal remodeling, and cell migration. MLK3 activity is linked to signaling networks downstream of small GTPases and receptor pathways that shape cellular stress responses and innate immune outputs. Dysregulated MAP3K11/MLK3 signaling has been implicated in pathological contexts involving aberrant MAPK activation, including oncogenic signaling adaptations and neuroinflammatory or neurodegenerative processes.

MLK3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP3K11 expression without altering the underlying DNA sequence.

MLK3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP3K11 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 MAP3K11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MLK3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP3K11 locus and enabling the study of MLK3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MLK3 pathway restoration in tumor cells with silenced or reduced MAP3K11 expression.

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