GIPC CRISPR Activation Plasmid (h): sc-402863-ACT

Human GIPC1 encodes GIPC (GAIP-interacting protein, C terminus), a PDZ domain–containing adaptor that binds the C-terminal motifs of diverse transmembrane receptors and trafficking components to coordinate endocytosis, recycling, and signal compartmentalization. GIPC links receptor complexes to the actin cytoskeleton and motor machinery through interactions with proteins such as myosin VI, shaping receptor availability and downstream signaling outputs. Through these scaffolding functions, GIPC1 has been implicated in pathways governing cell adhesion, migration, and growth factor receptor signaling dynamics. Dysregulated GIPC1 expression or altered GIPC-centered trafficking networks has been associated with phenotypes relevant to oncogenic signaling, invasive behavior, and other disorders where receptor turnover and spatial signaling control are perturbed.

GIPC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GIPC1 expression without altering the underlying DNA sequence.

GIPC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GIPC1 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 GIPC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GIPC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GIPC1 locus and enabling the study of GIPC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GIPC pathway restoration in tumor cells with silenced or reduced GIPC1 expression.

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