FOXL2 CRISPR Activation Plasmid (h): sc-403351-ACT

FOXL2 encodes a forkhead family transcription factor that plays a central role in ovarian development and granulosa cell identity by regulating lineage-specific gene expression programs. It modulates transcriptional networks controlling steroidogenesis, folliculogenesis, and cell-cycle balance through interactions with nuclear receptor signaling and chromatin-associated regulators. FOXL2 activity contributes to maintenance of ovarian function and cellular differentiation states, and dysregulation of FOXL2-dependent transcriptional control has been linked to reproductive endocrine disorders and tumor-associated gene expression signatures. As a DNA-binding regulator, FOXL2 is frequently studied for its effects on promoter/enhancer occupancy, epigenetic state, and context-dependent transcriptional output.

FOXL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXL2 expression without altering the underlying DNA sequence.

FOXL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXL2 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 FOXL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXL2 locus and enabling the study of FOXL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXL2 pathway restoration in tumor cells with silenced or reduced FOXL2 expression.

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