ERCC1 CRISPR Activation Plasmid (h): sc-400630-ACT

ERCC1 encodes a structure-specific endonuclease that forms a functional heterodimer with XPF (ERCC4) to execute incision steps during nucleotide excision repair (NER) and to resolve DNA interstrand crosslinks in coordination with the Fanconi anemia pathway. Through these activities, ERCC1 helps maintain genome stability, supports replication fork recovery, and limits the accumulation of DNA lesions caused by ultraviolet light and genotoxic agents. Altered ERCC1 function or expression is associated with impaired DNA repair capacity, elevated mutational burden, and cellular hypersensitivity to DNA-damaging stress. ERCC1-dependent repair processes are therefore widely studied in contexts such as carcinogenesis, aging-related genome maintenance, and mechanisms of resistance or susceptibility to DNA damage in experimental systems.

ERCC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ERCC1 expression without altering the underlying DNA sequence.

ERCC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ERCC1 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 ERCC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ERCC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ERCC1 locus and enabling the study of ERCC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ERCC1 pathway restoration in tumor cells with silenced or reduced ERCC1 expression.

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