c-Fgr CRISPR Activation Plasmid (m): sc-420347-ACT

Mouse Fgr encodes c-Fgr, a Src family non-receptor tyrosine kinase enriched in myeloid lineages that transduces signals downstream of immunoreceptors, integrins, and cytokine inputs to coordinate innate immune responses. c-Fgr participates in phosphorylation cascades that regulate actin cytoskeleton remodeling, cell adhesion and migration, phagocytosis, and reactive oxygen species production, integrating with pathways such as Fc receptor signaling and inflammatory kinase networks. Dysregulated Src family kinase activity, including altered Fgr signaling, has been linked to aberrant leukocyte activation and inflammatory phenotypes, making Fgr a useful node for studying immune homeostasis and myeloid cell function. In cancer biology, Fgr expression and signaling are also investigated for roles in tumor-associated myeloid cells and microenvironmental signaling dynamics.

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

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

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