PLC γ2 CRISPR Activation Plasmid (h): sc-400451-ACT

PLCG2 encodes phospholipase C gamma 2 (PLCγ2), a signal transduction enzyme that hydrolyzes PIP2 to generate IP3 and DAG, triggering intracellular Ca2+ mobilization and protein kinase C activation. PLCγ2 functions downstream of immunoreceptor and Fc receptor signaling and integrates cues from B-cell receptor pathways, innate immune receptors, and tyrosine kinase networks to regulate proliferation, differentiation, cytokine production, and phagocyte responses. In human biology, altered PLCG2 activity has been linked to immune dysregulation and inflammatory phenotypes, and it is widely studied in hematopoietic cells where calcium-dependent signaling programs shape immune cell fate. These properties make PLCG2 a useful node for dissecting receptor-proximal signaling circuits, transcriptional responses, and stimulus-dependent second messenger dynamics.

PLC γ2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLCG2 expression without altering the underlying DNA sequence.

PLC γ2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLCG2 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 PLCG2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLC γ2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLCG2 locus and enabling the study of PLC γ2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLC γ2 pathway restoration in tumor cells with silenced or reduced PLCG2 expression.

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