p39 CRISPR Activation Plasmid (h): sc-403667-ACT

Human CDK5R2 encodes p39, a neuronal regulatory subunit that activates cyclin-dependent kinase 5 (CDK5) and helps direct its subcellular localization and substrate specificity. The CDK5–p39 complex contributes to neuronal differentiation, cytoskeletal remodeling, neurite outgrowth, synaptic function, and activity-dependent signaling through phosphorylation networks that intersect with MAPK and calcium-regulated pathways. p39 is enriched in the nervous system and supports coordinated control of microtubule dynamics and vesicle trafficking during brain development and plasticity. Dysregulated CDK5 cofactor balance, including altered CDK5R2 expression or processing, has been associated with neurodegenerative and neurodevelopmental disease mechanisms in research contexts.

p39 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDK5R2 expression without altering the underlying DNA sequence.

p39 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDK5R2 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 CDK5R2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p39 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDK5R2 locus and enabling the study of p39-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p39 pathway restoration in tumor cells with silenced or reduced CDK5R2 expression.

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