AHCYL1 CRISPR Activation Plasmid (h): sc-417109-ACT

Human AHCYL1 encodes adenosylhomocysteinase-like 1, a non-catalytic paralog of S-adenosylhomocysteine hydrolase that functions as a regulatory component linking transmethylation capacity to intracellular signaling. AHCYL1 has been implicated in modulation of inositol 1,4,5-trisphosphate receptor–dependent calcium dynamics and associated programs such as ER-associated signaling, metabolic adaptation, and stress responses. Through these connections, AHCYL1 expression can influence epigenetic and transcriptional outputs downstream of methylation and Ca²⁺-responsive pathways. Dysregulation of methylation balance and Ca²⁺ signaling is frequently observed in inflammatory, neurobiological, and oncogenic contexts, making AHCYL1 a useful node for mechanistic investigation in disease-relevant cell models.

AHCYL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AHCYL1 expression without altering the underlying DNA sequence.

AHCYL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AHCYL1 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 AHCYL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AHCYL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AHCYL1 locus and enabling the study of AHCYL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AHCYL1 pathway restoration in tumor cells with silenced or reduced AHCYL1 expression.

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