DDB1 CRISPR Activation Plasmid (h): sc-402067-ACT

Human DDB1 encodes DNA damage-binding protein 1, a core adaptor of the CUL4–DDB1 E3 ubiquitin ligase complexes that coordinate ubiquitin-dependent turnover of chromatin-associated factors. DDB1 participates in nucleotide excision repair and broader genome maintenance by coupling UV-damage recognition to chromatin remodeling, replication stress responses, and proteostasis at stalled forks. Through regulation of cell-cycle progression and DNA repair capacity, altered DDB1 activity can influence genomic instability and transcriptional programs linked to oncogenic transformation and other proliferative disorders. Its central position in ubiquitin signaling makes DDB1 a useful node for studying DNA repair pathway choice, replication-coupled chromatin dynamics, and stress-adaptive remodeling of the epigenome.

DDB1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DDB1 expression without altering the underlying DNA sequence.

DDB1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DDB1 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 DDB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DDB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DDB1 locus and enabling the study of DDB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DDB1 pathway restoration in tumor cells with silenced or reduced DDB1 expression.

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