HIP12 CRISPR Activation Plasmid (h): sc-405552-ACT

Human HIP1R encodes huntingtin interacting protein 1 related (HIP12), an endocytic adaptor that links clathrin-coated pits to the actin cytoskeleton and participates in receptor internalization and membrane trafficking. HIP12 contains domains that bind clathrin and F-actin, supporting vesicle formation, cargo sorting, and coordination of cytoskeletal dynamics. Through these functions, HIP12 contributes to pathways governing endocytosis, cell signaling attenuation, and cytoskeleton-dependent remodeling events. Altered regulation of endocytic adaptors such as HIP1R has been associated in the literature with perturbations in signaling networks and cellular homeostasis relevant to neurobiology and cancer-related phenotypes, making HIP12 a useful target for mechanistic studies.

HIP12 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HIP1R expression without altering the underlying DNA sequence.

HIP12 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HIP1R 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 HIP1R transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HIP12 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HIP1R locus and enabling the study of HIP12-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HIP12 pathway restoration in tumor cells with silenced or reduced HIP1R expression.

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