KXD1 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of HeLa cells carrying a targeted disruption of the KXD1 gene. This loss-of-function model is designed to study the role of KXD1, an accessory subunit of the LKB1-STRAD-MO25 heterotrimeric complex, in cellular signaling and metabolism. The polyclonal format provides a heterogeneous knockout pool suitable for population-level analyses without the clonal artifacts often associated with single-cell-derived knockouts. No specific knockout mechanism is guaranteed; the product delivers a genetically diverse set of cells with KXD1 gene disruption.
The host cell line, HeLa, is a widely used epithelial cervical adenocarcinoma cell line positive for human papillomavirus type 18 (HPV-18). These immortalized cells are a standard model for human cervical cancer biology and HPV-associated oncogenesis. HeLa cells offer robust growth characteristics, ease of transfection, and well-characterized signaling networks, making them an ideal chassis for studying tumor suppressor pathways and metabolic regulation. The HPV-18 status adds relevance for exploring viral-host interactions in cancer metabolism.
KXD1 is an accessory subunit of the LKB1-STRAD-MO25 heterotrimeric complex, which functions as a master kinase upstream of AMPK under low energy conditions (high AMP/ATP). LKB1 directly phosphorylates AMPK at Thr172, leading to phosphorylation of targets such as ACC, TSC2, and ULK1, thereby inhibiting mTORC1 and promoting catabolic pathways. KXD1 interacts with LKB1 (STK11), STRAD (STRADA/STRADB), and MO25 (CAB39) to stabilize the complex. Disruption of KXD1 may attenuate AMPK activation, resulting in elevated mTORC1 signaling via Raptor and Rheb, altered energy sensing, and disrupted cell polarity. Downstream, PGC1?? and AMPK-responsive transcription factors are affected, offering a window into LKB1-dependent and -independent signaling.
In HeLa cells, which naturally harbor HPV-18 oncoproteins that can modulate AMPK and mTOR pathways, KXD1 knockout further perturbs the metabolic and proliferative signaling balance. This model is particularly relevant for investigating how loss of LKB1 complex function contributes to cancer phenotypes, including unchecked proliferation, metabolic reprogramming, and loss of cell polarity. It also provides a platform for studying Peutz-Jeghers syndrome-related mechanisms, where LKB1 mutations are causative. Additionally, the interplay between viral oncogenes and host energy-sensing pathways can be examined, offering insights into viral hijacking of metabolism.
Applications include western blotting for AMPK Thr172 phosphorylation, immunoprecipitation of LKB1 complex components, metabolic flux assays (e.g., Seahorse), mTOR signaling analysis, and phenotypic tests such as migration, invasion, and soft agar colony formation. RT-qPCR can quantify AMPK-regulated gene expression. These cells support drug screening for AMPK activators and mTOR inhibitors. For more information, please contact Ascent Research.
