FOXN1 CRISPR Activation Plasmid (h): sc-401498-ACT

FOXN1 (forkhead box N1) encodes a forkhead-family transcription factor that is essential for thymic epithelial cell differentiation and maintenance of a functional thymic microenvironment supporting T cell development. In epithelial compartments, FOXN1 regulates programs controlling keratinocyte maturation and epithelial lineage specification, linking chromatin-regulated transcription to tissue homeostasis. Altered FOXN1 activity has been associated with immunodeficiency phenotypes driven by impaired thymopoiesis, as well as epithelial barrier and differentiation defects. As a lineage-defining regulator, FOXN1 is widely used to study transcriptional networks governing thymus organogenesis, epithelial–immune crosstalk, and developmental gene regulation.

FOXN1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXN1 expression without altering the underlying DNA sequence.

FOXN1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXN1 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 FOXN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXN1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXN1 locus and enabling the study of FOXN1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXN1 pathway restoration in tumor cells with silenced or reduced FOXN1 expression.

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