PIPK I β CRISPR Activation Plasmid (h): sc-403758-ACT

PIP5K1B encodes phosphatidylinositol-4-phosphate 5-kinase type I beta (PIPK I β), a lipid kinase that generates phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) at cellular membranes. PI(4,5)P2 is a central signaling lipid that regulates actin cytoskeleton remodeling, focal adhesion dynamics, vesicular trafficking, and phosphoinositide-dependent signal transduction through pathways such as PLC/IP3-DAG and PI3K-AKT. Through these processes, PIPK I β contributes to cell migration, endocytosis, and membrane organization, linking phosphoinositide homeostasis to context-dependent changes in proliferation and immune cell function. Dysregulated phosphoinositide signaling and cytoskeletal control are recurrent features across cancer biology, inflammatory states, and neurobiology, making PIP5K1B a useful node for mechanistic studies of membrane-proximal signaling.

PIPK I β CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIP5K1B expression without altering the underlying DNA sequence.

PIPK I β CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIP5K1B 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 PIP5K1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PIPK I β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIP5K1B locus and enabling the study of PIPK I β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PIPK I β pathway restoration in tumor cells with silenced or reduced PIP5K1B expression.

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