PLC β1 CRISPR Activation Plasmid (h): sc-401141-ACT

Human PLCB1 encodes phospholipase C β1 (PLCβ1), a key effector downstream of G protein–coupled receptors that couples Gαq/11 signaling to phosphoinositide turnover. Upon activation, PLCβ1 hydrolyzes PIP2 to generate IP3 and diacylglycerol, triggering intracellular Ca2+ release and PKC activation that influence secretion, excitability, and transcriptional programs. This signaling axis integrates with MAPK and calcium-dependent pathways to shape neuronal and immune cell responses, and altered PLCB1 activity has been associated with neuropsychiatric and neurological phenotypes as well as dysregulated cell signaling in disease-relevant contexts. Studying PLCB1 regulation helps clarify how receptor-driven second messenger dynamics control downstream gene expression and cellular state.

PLC β1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLCB1 expression without altering the underlying DNA sequence.

PLC β1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLCB1 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 PLCB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLC β1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLCB1 locus and enabling the study of PLC β1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLC β1 pathway restoration in tumor cells with silenced or reduced PLCB1 expression.

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