CDYL CRISPR Activation Plasmid (h): sc-406624-ACT

Human CDYL (chromodomain Y-like) is a chromatin-associated coregulator that binds methylated lysines on histones and contributes to epigenetic control of transcription. Through its chromodomain and crotonyl-CoA hydratase-like activity, CDYL can influence histone crotonylation and acetylation states, supporting heterochromatin formation, transcriptional repression, and developmental gene programs. CDYL participates in pathways governing neuronal differentiation, germ cell development, and DNA damage–linked chromatin remodeling, reflecting its role in maintaining genome-regulatory states. Dysregulated CDYL expression or function has been associated with altered transcriptional networks reported in neurodevelopmental phenotypes and cancer-related epigenetic reprogramming, making it relevant for mechanistic studies of chromatin-dependent gene regulation.

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

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

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