ANKRD22 CRISPR Activation Plasmid (h): sc-416874-ACT

ANKRD22 (ankyrin repeat domain-containing protein 22) is a human ankyrin-repeat protein implicated in protein–protein interaction networks that shape cellular signaling and transcriptional regulation. Although its mechanistic roles remain incompletely defined, ANKRD22 expression has been linked to context-dependent changes in cell proliferation, metabolic adaptation, and stress-response programs. Transcriptomic studies report altered ANKRD22 levels across multiple cancer types, supporting investigation of its contribution to oncogenic signaling, tumor microenvironment adaptation, and biomarker-associated phenotypes. As a putative regulatory node, ANKRD22 is of interest for dissecting pathway cross-talk affecting growth control and cellular homeostasis.

ANKRD22 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ANKRD22 expression without altering the underlying DNA sequence.

ANKRD22 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ANKRD22 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 ANKRD22 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ANKRD22 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ANKRD22 locus and enabling the study of ANKRD22-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ANKRD22 pathway restoration in tumor cells with silenced or reduced ANKRD22 expression.

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