CaMKI CRISPR Activation Plasmid (h): sc-401136-ACT

Human CAMK1 encodes CaMKI, a calcium/calmodulin-dependent serine/threonine kinase that translates intracellular Ca2+ signals into phosphorylation events controlling transcriptional programs, cytoskeletal dynamics, and cell-cycle progression. CaMKI integrates into Ca2+/CaM signaling networks and can couple neuronal activity to gene expression through downstream kinase cascades and CREB-linked pathways, supporting neurite outgrowth and synaptic plasticity-related processes. Beyond the nervous system, CAMK1 activity has been implicated in broader signaling contexts that influence proliferation and stress responses, making it relevant to mechanistic studies of signaling dysregulation observed across complex diseases. Altered Ca2+-dependent kinase signaling is frequently associated with neurodevelopmental and neurodegenerative phenotypes as well as cancer-associated pathway rewiring, positioning CAMK1 as a useful node for pathway interrogation.

CaMKI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAMK1 expression without altering the underlying DNA sequence.

CaMKI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAMK1 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 CAMK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CaMKI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAMK1 locus and enabling the study of CaMKI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CaMKI pathway restoration in tumor cells with silenced or reduced CAMK1 expression.

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