FOXJ1 CRISPR Activation Plasmid (h): sc-402226-ACT

Human FOXJ1 encodes a forkhead box transcription factor that functions as a master regulator of motile ciliogenesis, coordinating basal body docking, axonemal assembly, and the transcriptional program required for multiciliated cell differentiation. FOXJ1 activity interfaces with developmental signaling and epithelial differentiation pathways, including cilia-associated transcriptional networks that shape airway and ependymal cell identity. Dysregulated FOXJ1 expression or ciliary gene programs is linked to impaired mucociliary clearance and broader ciliopathy-related phenotypes, making it relevant to studies of respiratory epithelium, reproductive tract function, and cerebrospinal fluid homeostasis. As a nuclear DNA-binding protein, FOXJ1 provides a tractable node for interrogating how transcriptional control of cilia genes influences barrier function, fluid flow, and innate defense mechanisms.

FOXJ1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXJ1 expression without altering the underlying DNA sequence.

FOXJ1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXJ1 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 FOXJ1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXJ1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXJ1 locus and enabling the study of FOXJ1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXJ1 pathway restoration in tumor cells with silenced or reduced FOXJ1 expression.

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