ANKS6 CRISPR Activation Plasmid (h): sc-406150-ACT

ANKS6 (ankyrin repeat and sterile alpha motif domain containing 6) encodes a scaffold-like protein implicated in primary cilium biology and centrosome-associated processes that coordinate epithelial polarity and tissue morphogenesis. It has been linked to nephronophthisis-related ciliopathy mechanisms, including regulation of ciliary signaling pathways that integrate developmental cues and mechanosensory inputs. ANKS6 participates in protein–protein interaction networks via ankyrin repeats and SAM domain-mediated assembly, supporting investigation of ciliary trafficking, cell cycle coordination, and planar cell polarity. Dysregulation or loss of ANKS6 function is associated with renal developmental abnormalities and cystic kidney phenotypes, making it relevant for studies of ciliopathies and kidney disease biology.

ANKS6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ANKS6 expression without altering the underlying DNA sequence.

ANKS6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ANKS6 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 ANKS6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ANKS6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ANKS6 locus and enabling the study of ANKS6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ANKS6 pathway restoration in tumor cells with silenced or reduced ANKS6 expression.

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