DTWD2 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human lung adenocarcinoma cell line NCI-H1975, with targeted disruption of the DTWD2 gene. This polyclonal population serves as a loss-of-function model to study the uncharacterized DTWD2 protein, a putative tRNA modification enzyme harboring a DTW domain. The CRISPR/Cas9-mediated gene disruption abolishes DTWD2 expression, enabling functional studies in a disease-relevant lung adenocarcinoma background.
The parental NCI-H1975 line is a well-established model of non-small cell lung cancer (NSCLC), isolated from a female patient with lung adenocarcinoma. These cells carry EGFR L858R and T790M activating mutations, making them resistant to first-generation EGFR tyrosine kinase inhibitors (TKIs) and a key system for investigating T790M-mediated drug resistance. Their epithelial morphology and retention of oncogenic drivers render NCI-H1975 cells ideal for studying molecular mechanisms underlying EGFR-mutant lung adenocarcinoma.
DTWD2 is a poorly characterized protein predicted to participate in tRNA modification, a process essential for translational accuracy and RNA metabolism. The DTW domain suggests a role in catalyzing nucleotide modifications on tRNA substrates. Although upstream regulators, downstream targets, and interacting partners remain undefined, DTWD2 likely functions within the tRNA modification pathway, potentially coordinating with other modifying enzymes. Disrupting DTWD2 in NCI-H1975 cells provides a tool to elucidate its contributions to RNA metabolism and cellular homeostasis in cancer.
In the context of NCI-H1975 cells, loss of DTWD2 may uncover linkages between tRNA modification and EGFR-driven oncogenic signaling. Altered tRNA modifications can influence the translation of proteins involved in cell proliferation, survival, or drug response, potentially affecting the malignant phenotype. This knockout model allows investigation of whether DTWD2 impacts tumor maintenance or modulates sensitivity to EGFR-targeted therapies, offering insights into novel functional dependencies in NSCLC.
This polyclonal knockout cell population is suitable for functional assays including western blotting to confirm DTWD2 loss, RT-qPCR for transcriptional analysis, and Sanger sequencing for genotyping. Phenotypic studies can include proliferation and migration assays, as well as drug sensitivity testing with EGFR inhibitors like osimertinib. The role of DTWD2 in tRNA modification can be directly assessed using mass spectrometry-based tRNA modification analysis. For additional information, please contact Ascent Research.
