The KLHL36 Knockout A-549 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human KLHL36 gene in the A-549 lung adenocarcinoma epithelial cell line (Homo sapiens). This loss-of-function model enables systematic interrogation of KLHL36-dependent ubiquitin-proteasome and mTORC1 signaling regulation.
Derived from a 58-year-old Caucasian male, A-549 is an adherent human lung adenocarcinoma cell line that serves as a widely accepted model for type II alveolar epithelial cells. It is extensively employed in respiratory disease, cancer biology, and xenobiotic metabolism studies, offering a physiologically relevant platform for investigating oncogenic signaling, drug metabolism, and epithelial cell biology.
KLHL36 functions as a substrate adaptor for the Cullin3-RING E3 ubiquitin ligase (CUL3) complex, mediating polyubiquitination and proteasomal degradation of Raptor, an essential scaffold component of mTORC1. Under conditions of nutrient sufficiency or growth factor stimulation, KLHL36 activity restricts mTORC1 signaling by promoting Raptor degradation; conversely, loss of KLHL36 stabilizes Raptor, leading to hyperactivation of mTORC1 and its downstream effectors S6K1 and 4E-BP1. The KLHL36-CUL3-Raptor axis integrates inputs from upstream regulators such as amino acid availability and cellular energy stress, and controls downstream targets including ULK1 and autophagy-related proteins p62/SQSTM1 and LC3, thereby coordinating protein synthesis, proliferation, and autophagic flux.
In the A-549 lung adenocarcinoma background, KLHL36 knockout perturbs the delicate balance of mTORC1 activity, which is frequently dysregulated in non-small cell lung cancer (NSCLC). The resulting Raptor stabilization and enhanced mTORC1 signaling create a hyper-proliferative state that mirrors oncogenic mTORopathy phenotypes, making this polyclonal knockout population a valuable tool for dissecting mTOR-driven tumorigenesis and metabolic reprogramming. Moreover, the interplay between KLHL36-dependent Raptor turnover and autophagy regulation offers insights into stress adaptation mechanisms in cancer cells.
Researchers can utilize these CRISPR/Cas9-edited polyclonal knockout cells in a wide array of assays, including western blotting for phospho-S6K1 and phospho-4E-BP1 to assess mTORC1 activation, immunoprecipitation-based detection of Raptor ubiquitination, cell proliferation (EdU/MTT) and apoptosis (Annexin V) analyses, and drug sensitivity testing with mTOR inhibitors such as rapamycin analogs. Furthermore, RT-qPCR for KLHL36 and immunofluorescence for mTOR subcellular localization allow precise validation. This model is particularly suited for investigating ubiquitin-proteasome system function, autophagy modulation, and the efficacy of mTOR-targeted therapeutics in lung adenocarcinoma. For further technical information or custom requests, please contact Ascent Research.
