BF-1 CRISPR Activation Plasmid (h): sc-402933-ACT

FOXG1 encodes the human brain factor-1 (BF-1) transcription factor, a forkhead family regulator that controls telencephalic and cortical progenitor proliferation, neuronal differentiation timing, and regional patterning during neurodevelopment. BF-1 modulates gene expression programs linked to cell cycle control and neuronal lineage specification, integrating with signaling networks such as TGF-β/SMAD and WNT/β-catenin-dependent transcriptional outputs. Dysregulated FOXG1 dosage or function perturbs excitatory–inhibitory balance and synaptic development, and pathogenic variants are associated with neurodevelopmental disorders including FOXG1 syndrome and epilepsy-related phenotypes. In cellular models, FOXG1 serves as a key node for studying chromatin-driven transcriptional control, fate decisions, and maturation trajectories in neural stem cells and neurons.

BF-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXG1 expression without altering the underlying DNA sequence.

BF-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXG1 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 FOXG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXG1 locus and enabling the study of BF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BF-1 pathway restoration in tumor cells with silenced or reduced FOXG1 expression.

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