MK CRISPR Activation Plasmid (h): sc-402064-ACT

Human MDK encodes midkine (MK), a secreted heparin-binding growth factor that modulates cell proliferation, survival, migration, and angiogenic responses during development and tissue remodeling. MK signaling interfaces with receptor tyrosine kinase and proteoglycan-mediated pathways, influencing downstream MAPK/ERK, PI3K/AKT, and inflammatory transcriptional programs that shape extracellular matrix dynamics and cell–cell communication. Dysregulated MDK expression has been reported across diverse malignancies and in pathological inflammation, ischemic injury responses, and fibrotic remodeling, where it can impact tumor–stroma interactions and immune cell recruitment. As a context-dependent regulator of stress and repair biology, MDK is widely studied in models of oncogenesis, neurobiology, and vascular biology.

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

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

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