G_{α 11} CRISPR Activation Plasmid (h): sc-401653-ACT

GNA11 encodes the human heterotrimeric G protein alpha subunit Gα11, a key transducer of signals from Gq/11-coupled GPCRs to downstream effector pathways. Upon receptor activation, Gα11 stimulates phospholipase C-β, promoting IP3/DAG production, intracellular Ca2+ mobilization, and protein kinase C activation, with downstream effects on MAPK/ERK and transcriptional programs controlling proliferation, secretion, and differentiation. GNA11 signaling contributes to regulation of vascular tone, endocrine function, and neuronal excitability, and dysregulation of this axis has been linked to oncogenic signaling and altered cell-state control in multiple disease contexts. As a proximal node in GPCR signal transduction, Gα11 is frequently interrogated to dissect pathway bias, calcium-dependent responses, and cross-talk with growth factor signaling.

Gα 11 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GNA11 expression without altering the underlying DNA sequence.

Gα 11 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GNA11 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 GNA11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Gα 11 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GNA11 locus and enabling the study of Gα 11-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Gα 11 pathway restoration in tumor cells with silenced or reduced GNA11 expression.

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