The DTWD1 Knockout Huh-7 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of Huh-7 human hepatocellular carcinoma cells harboring a targeted disruption of the DTWD1 gene. This knockout model is generated using CRISPR/Cas9 ribonucleoprotein complexes designed to introduce loss-of-function mutations within the DTWD1 locus, resulting in a heterogeneous pool of edited cells suitable for downstream functional analyses. The polyclonal format preserves genetic diversity while ensuring robust ablation of DTWD1 protein expression, enabling researchers to investigate the gene??s role without the limitations of single?cell clonal isolation.
The Huh-7 cell line is a well?differentiated hepatocellular carcinoma model originally established from the liver tumor of a 57?year?old Japanese male. These cells retain many hepatocyte?like features, including expression of liver?specific enzymes and metabolic pathways, making them a physiologically relevant system for studying hepatic cancer biology. The wild-type Huh-7 line has been extensively characterized in liver cancer research, and its genetic background provides a consistent platform for investigating tumor?associated gene functions.
DTWD1 encodes a protein containing a DTW domain, a conserved structural motif implicated in tRNA modification and translational control. Although the precise molecular function of DTWD1 remains under investigation, its DTW domain suggests a role in the biogenesis or function of modified nucleosides within tRNA, potentially regulating translation fidelity and protein synthesis rates. DTWD1 may interact with canonical tRNA modification enzymes, thereby influencing the cellular translation machinery. Disruption of DTWD1 in Huh-7 cells is expected to perturb these processes, leading to altered protein expression profiles that could affect cell proliferation, stress responses, and oncogenic signaling.
In the context of hepatocellular carcinoma, aberrant protein synthesis driven by dysregulated tRNA modifications has been linked to tumor progression and therapy resistance. The DTWD1 knockout Huh-7 model offers a valuable tool for dissecting how deficits in tRNA modification pathways contribute to liver cancer cell biology. By coupling this model with assays for proliferation, apoptosis, and metabolic flux, researchers can define the functional consequences of DTWD1 loss and its impact on tumor cell fitness, migration, and drug sensitivity.
This polyclonal knockout cell population is ideally suited for functional characterization of DTWD1 in hepatocellular carcinoma, mechanistic studies of tRNA modification pathways, and identification of novel therapeutic targets. Typical assays with these cells include western blotting and RT?qPCR for validation, proliferation (MTT/CCK?8) and colony formation for growth assessment, Transwell migration/invasion, apoptosis by Annexin V/PI flow cytometry, RNA?seq for transcriptome profiling, sorafenib drug sensitivity testing, and Seahorse metabolic flux analysis. For further information or to request a technical consultation, please contact Ascent Research.
