FGF-1 CRISPR Activation Plasmid (m): sc-420323-ACT

Mouse Fgf1 encodes fibroblast growth factor 1 (FGF-1), a pleiotropic heparin-binding growth factor that signals primarily through FGFR tyrosine kinase receptors to regulate cell proliferation, survival, migration, and differentiation. FGF-1 contributes to angiogenic and tissue remodeling programs and interfaces with MAPK/ERK, PI3K–AKT, and PLCγ signaling cascades, influencing transcriptional responses and cytoskeletal dynamics. In experimental models, altered FGF-1 activity has been linked to dysregulated wound repair, vascular and metabolic phenotypes, and aberrant growth signaling relevant to tumor biology and inflammation. These properties make Fgf1 a useful node for interrogating growth factor–driven signaling networks, extracellular matrix interactions, and context-dependent cellular plasticity.

FGF-1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Fgf1 expression without altering the underlying DNA sequence.

FGF-1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Fgf1 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 Fgf1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FGF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Fgf1 locus and enabling the study of FGF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FGF-1 pathway restoration in tumor cells with silenced or reduced Fgf1 expression.

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