FGF-2 CRISPR Activation Plasmid (h): sc-400324-ACT

Human FGF2 encodes fibroblast growth factor 2 (FGF-2), a heparin-binding mitogen that regulates cell proliferation, survival, migration, and differentiation across mesenchymal and neural lineages. FGF-2 signals primarily through FGFR tyrosine kinases to engage MAPK/ERK, PI3K/AKT, PLCγ, and STAT-associated programs, coordinating angiogenesis, tissue remodeling, and extracellular matrix dynamics. Dysregulated FGF2 expression or FGFR pathway activity is implicated in aberrant vascularization, fibrosis, and oncogenic signaling networks, and is frequently studied in contexts such as tumor–stroma interactions and neuroinflammation. As a pleiotropic growth factor, FGF-2 provides a tractable axis for interrogating receptor crosstalk, transcriptional state changes, and paracrine signaling in complex culture systems.

FGF-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FGF2 expression without altering the underlying DNA sequence.

FGF-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FGF2 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 FGF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FGF-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FGF2 locus and enabling the study of FGF-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FGF-2 pathway restoration in tumor cells with silenced or reduced FGF2 expression.

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