The BAG5 Knockout KYSE-150 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human esophageal squamous cell carcinoma line KYSE-150, with targeted disruption of the BAG5 gene. BAG5 encodes a co-chaperone that modulates Hsp70/Hsc70 activity and inhibits the E3 ubiquitin ligase Parkin, thereby suppressing protein degradation and mitophagy. This polyclonal pool contains a mixture of edited alleles, suitable for pooled functional assays without clonal selection.
The parental KYSE-150 cell line originates from a poorly differentiated esophageal squamous cell carcinoma from a Japanese patient. These cells express wild-type p53 and are tumorigenic, providing a relevant model for esophageal cancer biology. KYSE-150 cells exhibit malignant epithelial characteristics, including dysregulated proliferation and apoptosis, and are commonly employed to study oncogenic mechanisms and therapeutic responses in esophageal squamous cell carcinoma.
BAG5 functions as a co-chaperone that interacts with Hsp70 and Hsc70 and inhibits Parkin E3 ligase activity, blocking Parkin-mediated ubiquitination and proteasomal degradation. Through these interactions, BAG5 suppresses mitophagy and promotes cell survival under stress. The BAG5 interaction network includes Hsp70, Hsc70, Parkin, CHIP, and Bcl-2, linking protein quality control to apoptosis regulation. BAG5 expression is induced by HSF1 in response to cellular stress, and its downstream effects converge on Hsp70/Hsc70 chaperone modulation and Parkin inhibition. Key pathway components include BAG5, Hsp70, Parkin, ubiquitin, and CHIP.
In esophageal squamous cell carcinoma, apoptosis evasion and proteotoxic stress management are critical for tumor progression and drug resistance. BAG5 overexpression in KYSE-150 cells may confer a survival advantage by suppressing Parkin-dependent mitophagy and promoting anti-apoptotic signaling. Disruption of BAG5 in this wild-type p53 background allows dissection of BAG5-dependent survival mechanisms without confounding p53 mutations, offering a disease-relevant platform to study chaperone-mediated oncogenic pathways and chemoresistance.
These polyclonal knockout cells are suited for diverse applications: co-immunoprecipitation to assess Hsp70/Parkin complex alterations, Annexin V flow cytometry for apoptosis quantification, cell viability assays for drug sensitivity testing, and ubiquitination or mitophagy assays to monitor protein turnover. Genotyping PCR and RT-qPCR can confirm BAG5 disruption, while immunofluorescence enables subcellular study of chaperone networks. The model supports inhibitor screening and mechanistic studies of BAG5 in cancer biology. For further information, please contact Ascent Research.
