GFRα-3 CRISPR Activation Plasmid (h): sc-405061-ACT

GFRA3 encodes the human glycosylphosphatidylinositol-anchored receptor GFRα-3, a co-receptor that binds members of the glial cell line–derived neurotrophic factor (GDNF) ligand family and couples signaling to the RET receptor tyrosine kinase. Through RET-dependent activation of MAPK/ERK, PI3K/AKT, and related survival and differentiation pathways, GFRα-3 contributes to neuronal development, axon guidance, and maintenance of peripheral sensory and autonomic neurons. GFRA3 expression and signaling have been investigated in contexts of neurodevelopmental regulation, pain-related sensory neuron biology, and oncogenic pathway modulation in select tumor models where RET axis activity is implicated. As a cell-surface receptor component, it is also useful for studying ligand–receptor specificity, receptor complex assembly, and downstream transcriptional responses.

GFRα-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GFRA3 expression without altering the underlying DNA sequence.

GFRα-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GFRA3 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 GFRA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GFRα-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GFRA3 locus and enabling the study of GFRα-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GFRα-3 pathway restoration in tumor cells with silenced or reduced GFRA3 expression.

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