The ABHD14B Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the human ABHD14B gene has been disrupted. This loss-of-function model enables the study of ABHD14B-mediated lysine deacetylation and its role in metabolic regulation. The polyclonal nature of the knockout cell population provides a heterogeneous genetic background, capturing a range of editing outcomes across the cell pool, which is suitable for initial functional screening and pathway analysis.
The parental HEK293T cell line is a human embryonic kidney epithelial cell line stably expressing the SV40 large T antigen, derived from the HEK293 lineage. This widely employed host exhibits high transfection efficiency, robust recombinant protein expression capacity, and neomycin resistance, making it a versatile platform for viral production, gene function studies, and metabolic investigations. Its adherent growth and well-characterized biology facilitate reproducible experimental workflows.
ABHD14B encodes a lysine deacetylase that specifically removes acetyl groups from oxyglutarate dehydrogenase (OGDH), a key enzyme of the tricarboxylic acid (TCA) cycle. This deacetylation event modulates OGDH activity, thereby influencing TCA cycle flux and cellular respiration. The functional network includes upstream metabolic inputs such as acetyl-CoA and NAD+ levels, direct interaction with the OGDH complex (comprising OGDH, DLST, and DLD), and downstream effects on alpha-ketoglutarate metabolism and TCA cycle intermediates like citrate and isocitrate. Loss of ABHD14B is predicted to result in OGDH hyperacetylation, potentially altering metabolic flux and contributing to metabolic dysregulation.
In the HEK293T context, disruption of ABHD14B provides a valuable system to dissect the consequences of impaired TCA cycle deacetylation. Given the high metabolic activity of HEK293T cells and their common use in cancer metabolism research, this knockout model is particularly relevant for exploring metabolic rewiring mechanisms. The absence of ABHD14B may mimic states of aberrant protein acetylation observed in certain cancers and metabolic disorders, allowing researchers to interrogate how deacetylation-dependent control of OGDH impacts energy production and biosynthesis.
Researchers can employ the ABHD14B Knockout HEK293T Polyclonal Cells in a variety of assays, including western blotting to assess global and OGDH-specific acetylation changes, real-time metabolic flux analysis using Seahorse platforms, and co-immunoprecipitation to examine interactions within the OGDH complex. Additional applications include RT-qPCR profiling of metabolic gene expression, OGDH activity measurements, and immunofluorescence studies to monitor subcellular localization. This knockout population is a starting point for further clonal isolation if desired. For detailed technical specifications or customized support, please contact Ascent Research.
