PICK1 CRISPR Activation Plasmid (h): sc-403683-ACT

Human PICK1 (protein interacting with C kinase 1) is a PDZ and BAR domain–containing scaffold that couples membrane curvature sensing to receptor trafficking and signal transduction. PICK1 regulates endocytosis, recycling, and synaptic localization of diverse membrane proteins, including AMPA-type glutamate receptors and transporters, influencing synaptic plasticity and intracellular kinase signaling. Through its roles in vesicular trafficking and cytoskeletal organization, PICK1 participates in neuronal communication and polarized transport processes in multiple cell types. Dysregulated PICK1-associated pathways have been investigated in the context of neurological dysfunction and altered receptor homeostasis, making it a useful node for studying membrane-proximal signaling networks.

PICK1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PICK1 expression without altering the underlying DNA sequence.

PICK1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PICK1 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 PICK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PICK1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PICK1 locus and enabling the study of PICK1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PICK1 pathway restoration in tumor cells with silenced or reduced PICK1 expression.

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