PLC β2 CRISPR Activation Plasmid (h): sc-400984-ACT

Human PLCB2 encodes phospholipase C beta 2 (PLCβ2), a phosphoinositide-specific phospholipase that couples activated GPCR signaling to second messenger production. PLCβ2 hydrolyzes PIP2 to generate IP3 and DAG, promoting intracellular Ca2+ mobilization and PKC activation that regulate chemotaxis, degranulation, cytokine production, and other effector functions. This pathway integrates with PI3K, MAPK, and small GTPase signaling to shape cytoskeletal remodeling and transcriptional programs, particularly in hematopoietic and immune cell contexts. Dysregulated PLCB2 activity and Ca2+-dependent signaling has been associated with altered inflammatory responses and oncogenic signaling networks, supporting its utility as a mechanistic node for pathway interrogation.

PLC β2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLCB2 expression without altering the underlying DNA sequence.

PLC β2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLCB2 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 PLCB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLC β2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLCB2 locus and enabling the study of PLC β2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLC β2 pathway restoration in tumor cells with silenced or reduced PLCB2 expression.

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