ANKRD9 CRISPR Activation Plasmid (m): sc-428809-ACT

Mouse Ankrd9 encodes ANKRD9, an ankyrin repeat–containing protein implicated in protein–protein interaction networks that shape intracellular signaling and cell state regulation. Although the molecular function of ANKRD9 remains incompletely characterized, ankyrin repeat proteins commonly modulate adaptor/scaffold activity, influencing pathways linked to cytoskeletal organization, vesicular trafficking, and transcriptional responses to metabolic and stress cues. Ankrd9 expression has been explored in contexts relevant to tissue homeostasis and differentiation, where shifts in regulatory networks can contribute to phenotypes associated with inflammation, metabolic imbalance, and dysregulated growth programs. As a result, Ankrd9 serves as a useful node for probing pathway connectivity and gene network remodeling in mouse cellular and in vivo model systems.

ANKRD9 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ankrd9 expression without altering the underlying DNA sequence.

ANKRD9 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ankrd9 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 Ankrd9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ANKRD9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ankrd9 locus and enabling the study of ANKRD9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ANKRD9 pathway restoration in tumor cells with silenced or reduced Ankrd9 expression.

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