GFAP CRISPR Activation Plasmid (h): sc-400130-ACT

Glial fibrillary acidic protein (GFAP) is an intermediate filament protein that defines astrocyte identity and contributes to cytoskeletal organization, cellular mechanical stability, and process remodeling in the central nervous system. GFAP dynamics intersect with astrocyte reactivity programs and signaling networks such as JAK/STAT and MAPK that modulate gliosis, neuroinflammation, and tissue repair responses. Altered GFAP expression and filament architecture are linked to astrocyte dysfunction in neurological and neurodegenerative conditions, and GFAP is widely used as a molecular readout for astrocyte activation state and lineage. Experimental modulation of GFAP supports studies of astrocyte maturation, neuron–glia interactions, and stress-responsive cytoskeletal remodeling.

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

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

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