Galanin CRISPR Activation Plasmid (h): sc-402730-ACT

Human GAL encodes galanin, a secreted neuropeptide that signals through G protein–coupled receptors (GALR1–3) to regulate neuronal excitability, neurotransmitter release, and neuroendocrine function. Galanin modulates second-messenger pathways including cAMP/PKA, MAPK/ERK, and calcium signaling, influencing processes such as appetite regulation, nociception, stress responsiveness, and autonomic control. In the nervous system and peripheral tissues, GAL activity contributes to synaptic plasticity and neuroimmune crosstalk, making it relevant to mechanistic studies of neurological and metabolic phenotypes. Altered galanin signaling has been associated in the literature with disorders involving pain processing, mood regulation, seizure susceptibility, and energy homeostasis, supporting its use as a pathway node in disease-relevant models.

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

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

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