AHDC1 CRISPR Activation Plasmid (h): sc-408863-ACT

AHDC1 (AT-hook DNA binding motif containing 1) encodes a nuclear chromatin-associated protein implicated in transcriptional regulation and higher-order genome organization through DNA binding and interaction with regulatory complexes. It is linked to processes that govern neurodevelopment, neuronal differentiation, and maintenance of gene expression programs, consistent with roles in chromatin remodeling and RNA-associated regulatory networks. Genetic disruption of AHDC1 is associated with Xia-Gibbs syndrome, a neurodevelopmental disorder characterized by developmental delay and neurological phenotypes, highlighting its importance in brain-relevant transcriptional control. Altered AHDC1 function is therefore of interest for studying epigenetic regulation, neuronal gene networks, and genotype–phenotype relationships in human cellular models.

AHDC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AHDC1 expression without altering the underlying DNA sequence.

AHDC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AHDC1 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 AHDC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AHDC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AHDC1 locus and enabling the study of AHDC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AHDC1 pathway restoration in tumor cells with silenced or reduced AHDC1 expression.

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