Fer CRISPR Activation Plasmid (h): sc-402734-ACT

Human FER encodes Fer, a non-receptor protein tyrosine kinase in the FES/Fer family that transduces signals downstream of growth factor receptors and adhesion complexes. Fer participates in phosphorylation-dependent control of cytoskeletal dynamics, cell–cell junction integrity, and cell motility through pathways involving integrin signaling, cadherin-associated complexes, and Rho family GTPase regulation. By modulating MAPK and PI3K/AKT-associated signaling outputs, Fer helps coordinate proliferation and survival responses to extracellular cues. Dysregulated FER activity or expression has been linked to altered invasive behavior and aberrant signaling networks in multiple cancer-relevant contexts, supporting its use as a mechanistic node in studies of oncogenic signaling and metastasis-associated phenotypes.

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

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

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