The HLTF Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from NCI-H1975 human lung adenocarcinoma cells. This polyclonal format, generated by CRISPR/Cas9-mediated disruption of the HLTF gene, provides a heterogeneous editing pool without clonal selection, enabling population-level loss-of-function studies in an isogenic background. It is designed for investigating HLTF-dependent DNA repair, replication stress, and tumor suppression in a non-small cell lung cancer model.
NCI-H1975 is a human lung adenocarcinoma cell line with EGFR L858R/T790M mutations, widely used as an NSCLC model for oncogenic signaling and therapeutic resistance. These epithelial cells retain molecular features of lung adenocarcinoma, providing a clinically relevant host for gene-edited variants. The HLTF knockout in this background allows dissection of DNA repair?CEGFR crosstalk in cancer biology.
HLTF encodes a SWI/SNF-family ATP-dependent DNA helicase that functions as a tumor suppressor through post-replication repair and fork reversal. It is activated by ATM/ATR kinases upon DNA damage and interacts with PCNA, RAD51, SHPRH, and MMS22L to facilitate homologous recombination and lesion bypass. HLTF also transcriptionally regulates repair factors and chromatin remodelers. Knockout disrupts these processes, causing DNA repair deficiency and genome instability.
In EGFR-mutant NCI-H1975 cells, HLTF loss exacerbates replication stress and sensitizes cells to genotoxic agents like cisplatin and PARP inhibitors. This model is valuable for studying synthetic lethality and drug response profiling. It also enables exploration of how HLTF inactivation cooperates with oncogenic EGFR to drive tumor progression, and investigation of epigenetic HLTF silencing seen in colorectal and lung cancers.
Research applications encompass DNA repair mechanism studies, replication stress biology, and tumor suppressor research. Common assays include western blotting, RT-qPCR, ??-H2AX/RAD51 immunofluorescence, cell cycle and apoptosis flow cytometry, co-immunoprecipitation, and drug sensitivity testing with cisplatin and PARP inhibitors. Additional techniques such as comet assays and migration/invasion experiments extend utility. For detailed protocols, validation, or ordering information, please contact Ascent Research.
