GFRαL CRISPR Activation Plasmid (h): sc-415831-ACT

Human GFRAL (GDNF family receptor alpha-like) encodes GFRαL, a GPI-anchored co-receptor that confers ligand specificity to RET signaling in response to GDF15. This receptor complex is enriched in hindbrain neurons and links extracellular cues to intracellular kinase cascades such as MAPK/ERK and PI3K/AKT, shaping neuronal activity and transcriptional programs involved in systemic energy balance. Altered GDF15–GFRαL–RET axis activity has been associated with metabolic dysregulation and cachexia-related biology, making GFRAL a useful entry point for dissecting neuroendocrine control of appetite and stress-associated weight change. In vitro, modulation of GFRAL expression supports studies of receptor trafficking, ligand-dependent signaling, and downstream gene networks in relevant neuronal or engineered cell models.

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

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

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