PLC γ1 CRISPR Activation Plasmid (h): sc-400472-ACT

PLCG1 encodes phospholipase C gamma 1 (PLCγ1), a receptor-proximal signaling enzyme activated downstream of receptor tyrosine kinases and immune receptors. Upon activation, PLCγ1 hydrolyzes PIP2 to generate IP3 and DAG, triggering intracellular Ca2+ mobilization and PKC activation that coordinate transcriptional programs, cytoskeletal remodeling, and cell migration. This signaling node integrates inputs from pathways such as EGFR/PDGFR/FGFR and T cell receptor signaling, shaping proliferation and differentiation responses. Dysregulated PLCG1 activity and pathway rewiring have been linked to aberrant growth factor signaling and immune-related pathologies, making it a useful target for mechanistic studies of phosphoinositide-driven signal transduction.

PLC γ1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLCG1 expression without altering the underlying DNA sequence.

PLC γ1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLCG1 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 PLCG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLC γ1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLCG1 locus and enabling the study of PLC γ1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLC γ1 pathway restoration in tumor cells with silenced or reduced PLCG1 expression.

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