TFIIH p80 CRISPR Activation Plasmid (h): sc-402231-ACT

ERCC2 encodes the human TFIIH p80 helicase (XPD), an essential subunit of the TFIIH complex that couples ATP-dependent DNA unwinding to transcription initiation by RNA polymerase II and nucleotide excision repair (NER). Through its roles in transcription-coupled repair and global genome NER, TFIIH p80 supports genome stability by enabling recognition and removal of bulky DNA lesions such as UV-induced photoproducts. ERCC2 also influences cell-cycle progression and the cellular DNA damage response through TFIIH-mediated coordination of repair and transcriptional programs. Pathogenic ERCC2 variants are associated with DNA repair disorders including xeroderma pigmentosum, trichothiodystrophy, and Cockayne syndrome, making ERCC2 a widely used locus for studying NER dysfunction and transcription-repair coupling.

TFIIH p80 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ERCC2 expression without altering the underlying DNA sequence.

TFIIH p80 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ERCC2 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 ERCC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TFIIH p80 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ERCC2 locus and enabling the study of TFIIH p80-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TFIIH p80 pathway restoration in tumor cells with silenced or reduced ERCC2 expression.

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