FGFR-5 CRISPR Activation Plasmid (m): sc-431146-ACT

Mouse Fgfrl1 encodes FGFR-5, an atypical fibroblast growth factor receptor–like protein that can modulate FGF signaling outputs by influencing ligand availability and receptor complex dynamics. FGFR-5 is implicated in the regulation of developmental processes including organogenesis, tissue patterning, and cell–cell interactions, with downstream effects on pathways commonly linked to FGFR biology such as MAPK/ERK and PI3K/AKT signaling. In mammalian systems, altered FGFRL1 function has been associated with defects in morphogenesis and cardiopulmonary development, supporting its relevance to congenital disease mechanisms. As a noncanonical receptor family member, FGFR-5 provides a useful entry point for dissecting how FGF pathway tuning affects differentiation and tissue homeostasis.

FGFR-5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Fgfrl1 expression without altering the underlying DNA sequence.

FGFR-5 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Fgfrl1 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 Fgfrl1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FGFR-5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Fgfrl1 locus and enabling the study of FGFR-5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FGFR-5 pathway restoration in tumor cells with silenced or reduced Fgfrl1 expression.

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