Fra2 CRISPR Activation Plasmid (h): sc-400250-ACT

Human FOSL2 encodes Fra2 (FOS-like antigen 2), a bZIP transcription factor that heterodimerizes with JUN proteins to form AP-1 complexes regulating stimulus-responsive gene expression. Fra2 integrates MAPK/ERK and JNK signaling to control programs linked to cell-cycle progression, differentiation, extracellular matrix remodeling, and inflammatory responses. Dysregulated FOSL2/AP-1 activity has been associated with oncogenic transcriptional states, tissue fibrosis, and chronic inflammation, making it a useful node for dissecting transcriptional control of stress and growth factor pathways. Fra2-dependent regulatory networks are commonly studied in contexts such as epithelial–mesenchymal transition, stromal activation, and cytokine-driven signaling.

Fra2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOSL2 expression without altering the underlying DNA sequence.

Fra2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOSL2 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 FOSL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fra2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOSL2 locus and enabling the study of Fra2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fra2 pathway restoration in tumor cells with silenced or reduced FOSL2 expression.

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