The HTD2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to study the mitochondrial enoyl-CoA reductase HTD2. This product provides a mixed population of HeLa cells carrying targeted disruptions in the HTD2 gene, enabling loss-of-function analysis in a heterogeneous cell pool. The polyclonal format allows researchers to assess gene function in the context of a diverse genetic background, which can better model natural variation compared to clonal isolates. These cells are suited for biochemical, metabolic, and functional assays requiring HTD2-deficient HeLa derivatives.
The HeLa host cell line is a human cervical adenocarcinoma-derived epithelial cell line immortalized by human papillomavirus type 18 (HPV18). HeLa cells are aneuploid and have been widely adopted in cancer biology and molecular research because of their robust growth and ease of manipulation. Their well-characterized signaling networks and metabolic properties make them a standard model for studying mitochondrial function, oncogenesis, and drug responses. This host background provides a reproducible platform for investigating the consequences of HTD2 disruption in a cancerous cellular environment.
HTD2 encodes a mitochondrial enoyl-CoA reductase catalyzing the final step of mitochondrial fatty acid synthesis. This reaction converts trans-2-enoyl-ACP to saturated acyl-ACP, producing octanoyl-ACP, the precursor for lipoic acid. Lipoic acid is a critical cofactor for pyruvate dehydrogenase and ??-ketoglutarate dehydrogenase. HTD2 functions in a complex with OXSM, HSD17B8, and ACP, with LIAS completing lipoic acid synthesis. Expression of mtFAS genes is regulated by PGC-1??, NRF1, and TFAM under metabolic stress. Disruption of HTD2 impairs octanoyl-ACP production, reducing lipoic acid availability and compromising mitochondrial dehydrogenase activity, leading to respiratory dysfunction and energetic stress.
In HeLa cells, HTD2 knockout highlights the dependence of cancer cells on mitochondrial metabolism. Loss of HTD2 disrupts lipoic acid-dependent enzyme function, impairing oxidative phosphorylation and increasing ROS. This model helps dissect how mitochondrial fatty acid synthesis supports proliferation and survival under stress. Moreover, HTD2 dysfunction is linked to MEPAN syndrome and mitochondrial disorders involving neurodegeneration and optic atrophy, making these cells relevant for disease modeling.
Researchers can employ these polyclonal knockout cells in diverse experimental workflows. Metabolic flux can be assessed using Seahorse respirometry, while lipoic acid levels and mtFAS intermediate accumulation can be measured by targeted mass spectrometry. Western blotting for MECR and RT-qPCR for mtFAS-related transcripts allow confirmation of gene disruption. Additional assays include ROS detection, apoptosis analysis, and mitochondrial morphology immunofluorescence. These cells are suitable for investigating mitochondrial metabolism, neurodegeneration modeling, lipoic acid synthesis, cancer metabolism, and drug target validation. For additional information or technical support, please contact Ascent Research.
