Cdk5 CRISPR Activation Plasmid (m): sc-419602-ACT

Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine kinase that, unlike cell cycle CDKs, is primarily activated in post-mitotic cells through association with p35/p39 and regulates neuronal development and function. In mouse, Cdk5 controls cytoskeletal dynamics, axon guidance, synaptic vesicle trafficking, and activity-dependent signaling by phosphorylating substrates that modulate microtubule stability, endocytosis, and transcriptional responses. It intersects with pathways governing calcium signaling, kinase cascades, and proteostasis, integrating cues that shape neuronal morphology and network plasticity. Dysregulated Cdk5 activity has been linked to mechanisms relevant to neurodegeneration, neuroinflammation, and neuropsychiatric phenotypes, making it a frequent target for studies of neuronal stress responses and circuit-level dysfunction.

Cdk5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cdk5 expression without altering the underlying DNA sequence.

Cdk5 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cdk5 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 Cdk5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cdk5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cdk5 locus and enabling the study of Cdk5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cdk5 pathway restoration in tumor cells with silenced or reduced Cdk5 expression.

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