The GTF2H5 knockout Jurkat polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from Jurkat human T lymphocytes, providing a loss-of-function model for the GTF2H5 gene. GTF2H5 encodes a critical subunit of the transcription factor IIH (TFIIH) complex, essential for RNA polymerase II transcription initiation and nucleotide excision repair (NER). This polyclonal pool, created via CRISPR/Cas9-mediated gene disruption, yields heterogeneous edits that avoid clonal artifacts and capture population-level variation, enabling robust assessment of GTF2H5 function in a well-characterized T-cell background.
The Jurkat cell line, derived from acute T-cell leukemia, is an immortalized human T lymphocyte widely used to model T-cell receptor signaling, apoptosis, and immune function. Its extensive characterization and genetic tractability make it ideal for studying gene function in lymphoid cells, with well-defined signaling and apoptotic pathways that facilitate examination of DNA repair and cell survival crosstalk.
GTF2H5 (p8/TTD-A) is an integral TFIIH subunit that stabilizes the core complex composed of XPB, XPD, p62, p52, p44, and p34. It directly interacts with Cdk7, Cyclin H, and MAT1 to modulate kinase activity for RNA polymerase II recruitment and phosphorylation during transcription initiation. In NER, GTF2H5-anchored TFIIH unwinds damaged DNA, enabling lesion recognition and incision by factors like XPA, XPC, XPF-ERCC1, and XPG. Consequently, GTF2H5 functions upstream of RNA polymerase II-dependent gene transcription and downstream of TFIIH assembly factors, connecting it to cell cycle control and genome integrity.
In Jurkat T cells, GTF2H5 knockout destabilizes TFIIH, causing severe defects in transcription-coupled repair and global transcription, with heightened UV sensitivity and apoptosis. This mirrors trichothiodystrophy and photosensitivity disorders, where GTF2H5 mutations impair NER without full cancer proneness. The Jurkat model further permits investigation of how compromised transcription and repair impact T-cell activation, proliferation, and immune functions.
Applications span transcription-coupled repair analysis, trichothiodystrophy modeling, and DNA damage response studies, with assays such as Western blotting for GTF2H5 depletion, UV sensitivity and comet assays, RNA-seq and RT-qPCR for NER gene expression, flow cytometry for apoptosis, and immunofluorescence for TFIIH localization. In drug discovery, these cells enable screening of NER-targeted compounds and DNA-damaging agent sensitizers. For further technical inquiries or custom orders, please contact Ascent Research.
