TFIIH p62 CRISPR Activation Plasmid (h): sc-402239-ACT

GTF2H1 encodes the TFIIH p62 subunit, a core component of the TFIIH complex that coordinates RNA polymerase II transcription initiation with nucleotide excision repair (NER). TFIIH integrates promoter opening, transcription start site selection, and DNA damage recognition/repair through interactions with other TFIIH subunits and repair factors, linking basal transcription to genome maintenance. Altered TFIIH function and NER defects are associated with UV sensitivity and transcription-coupled repair syndromes, underscoring the importance of p62 for cellular responses to genotoxic stress. As a scaffold-like factor within TFIIH, TFIIH p62 provides a tractable node for dissecting cross-talk between transcriptional regulation and DNA repair pathways.

TFIIH p62 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GTF2H1 expression without altering the underlying DNA sequence.

TFIIH p62 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GTF2H1 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 GTF2H1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TFIIH p62 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GTF2H1 locus and enabling the study of TFIIH p62-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TFIIH p62 pathway restoration in tumor cells with silenced or reduced GTF2H1 expression.

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