Fyn CRISPR Activation Plasmid (m2): sc-420431-ACT-2

Mouse Fyn encodes a Src family non-receptor tyrosine kinase that functions as a proximal signaling hub downstream of immune receptors, integrins, and growth factor receptors, regulating phosphorylation-dependent control of cytoskeletal dynamics, cell adhesion, migration, and proliferation. In T cells and other hematopoietic lineages, Fyn contributes to TCR signaling and immune activation through pathways including MAPK/ERK, PI3K–AKT, and modulation of phosphatase and adaptor complexes, while in the nervous system it influences synaptic signaling and myelination-related processes. Dysregulated Fyn activity has been linked to altered inflammatory responses, neurodegeneration-associated signaling, and oncogenic signaling networks in model systems, making it relevant for studies of immune regulation, neuronal function, and tumor biology. Gene editing of mouse Fyn supports mechanistic dissection of kinase-dependent pathways, phosphorylation networks, and cell-type–specific phenotypes in vitro and in vivo.

Fyn CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Fyn expression without altering the underlying DNA sequence.

Fyn CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Fyn 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 Fyn transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fyn expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Fyn locus and enabling the study of Fyn-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fyn pathway restoration in tumor cells with silenced or reduced Fyn expression.

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