The CAB39L Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the CAB39L gene. This polyclonal format avoids clonal artifacts and represents a heterogeneous pool of gene-disrupted cells, enabling robust assessment of CAB39L-dependent phenotypes.
The HAP1 cell line is a near-haploid human line derived from KBM-7 chronic myeloid leukemia cells. Its near-haploid karyotype simplifies genetic analysis by minimizing redundant alleles, making it an ideal host for CRISPR-mediated knockout models. HAP1 cells maintain key signaling pathways relevant to cancer and metabolism, providing a clean genetic background for pathway dissection.
CAB39L encodes a scaffold protein that complexes with STE20-related adaptor alpha (STRAD??) and the tumor suppressor kinase LKB1 (STK11), enhancing LKB1-mediated phosphorylation of AMPK. By binding MO25 and stabilizing this heterotrimeric complex, CAB39L facilitates AMPK activation under energy stress, leading to downstream phosphorylation of TSC2 and inhibition of mTORC1. CAB39L also interacts with AMPK subunits and contributes to the regulation of MARK kinases and Hippo pathway components, thereby integrating metabolic cues with cell polarity and growth control. Consequently, loss of CAB39L disrupts AMPK signaling, resulting in impaired energy homeostasis and unchecked mTOR activity.
In HAP1 cells, CAB39L knockout provides a simplified model to study the AMPK?CmTOR axis and polarity without confounding diploid gene redundancy. The near-haploid state accentuates functional consequences of gene loss, enabling clearer delineation of CAB39L??s role in metabolic reprogramming and proliferation. This system is particularly valuable for investigating metabolic vulnerabilities in cancer and evaluating compounds that target the LKB1?CAMPK pathway.
Researchers can deploy this knockout population in diverse assays: Western blotting for phospho-AMPK (Thr172) and phospho-mTOR targets, RT?qPCR of AMPK-responsive genes, metabolic flux analysis, and proliferation assays. Drug sensitivity profiling with metformin or AICAR helps assess reliance on CAB39L for stress responses, while migration assays explore polarity defects. For additional technical information or to discuss custom applications, please contact Ascent Research.
