PLC β3 CRISPR Activation Plasmid (h): sc-401181-ACT

PLCB3 encodes phospholipase C beta 3 (PLCβ3), a key effector downstream of G protein–coupled receptors that hydrolyzes phosphatidylinositol 4,5-bisphosphate to generate inositol 1,4,5-trisphosphate and diacylglycerol. This signaling axis elevates intracellular Ca2+ and activates protein kinase C, integrating inputs that regulate secretion, migration, cytoskeletal remodeling, and transcriptional programs. PLCβ3 activity links GPCR signaling to phosphoinositide turnover and calcium-dependent pathways that shape innate and adaptive immune responses as well as broader cell-state transitions. Dysregulation of phospholipase C signaling has been associated with aberrant proliferation and inflammatory phenotypes, making PLCB3 a useful node for mechanistic studies of signal transduction and disease-relevant cellular behaviors.

PLC β3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLCB3 expression without altering the underlying DNA sequence.

PLC β3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLCB3 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 PLCB3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLC β3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLCB3 locus and enabling the study of PLC β3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLC β3 pathway restoration in tumor cells with silenced or reduced PLCB3 expression.

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