ANKFN1 CRISPR Activation Plasmid (h): sc-406947-ACT

Human ANKFN1 encodes an ankyrin repeat and fibronectin type III domain–containing protein thought to support protein–protein interactions and scaffolding functions that influence subcellular organization and signaling dynamics. Although the gene remains incompletely characterized, domain architecture suggests potential roles in coordinating cytoskeletal-associated processes and regulatory complexes that modulate cellular responses to stress and differentiation cues. Transcriptomic and genomic studies have reported altered ANKFN1 expression or association signals in select neurological and developmental phenotypes, highlighting it as a candidate for mechanistic investigation. Defining ANKFN1-dependent interaction networks and downstream transcriptional programs can inform pathway mapping in cell models relevant to tissue development and disease biology.

ANKFN1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ANKFN1 expression without altering the underlying DNA sequence.

ANKFN1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ANKFN1 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 ANKFN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ANKFN1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ANKFN1 locus and enabling the study of ANKFN1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ANKFN1 pathway restoration in tumor cells with silenced or reduced ANKFN1 expression.

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