Dyskerin CRISPR Activation Plasmid (h): sc-401278-ACT

Human DKC1 encodes dyskerin, an essential nucleolar protein that stabilizes the H/ACA small nucleolar RNP complex and catalyzes pseudouridylation of rRNA and snRNA, supporting ribosome biogenesis and pre-mRNA splicing fidelity. Dyskerin also associates with the telomerase holoenzyme through binding the telomerase RNA component, contributing to telomere maintenance and genome stability. Through these functions, DKC1 integrates RNA processing with cell cycle control, DNA damage responses, and proteostasis under proliferative stress. Dysregulation or mutation of DKC1 is linked to telomere biology disorders and impaired hematopoietic and epithelial homeostasis, making it a key target for mechanistic studies of ribonucleoprotein assembly and nucleolar function.

Dyskerin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DKC1 expression without altering the underlying DNA sequence.

Dyskerin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DKC1 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 DKC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dyskerin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DKC1 locus and enabling the study of Dyskerin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dyskerin pathway restoration in tumor cells with silenced or reduced DKC1 expression.

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