The BAG3 Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the BAG3 gene. This product provides a genetically disrupted pool of cells, enabling investigation of BAG3-dependent cellular processes without the confounding effects of clonal selection. The polyclonal format preserves population-level heterogeneity while ensuring robust target-gene disruption, making it suitable for pooled functional assays and screening applications.
The DLD-1 host cell line is a well-characterized human colorectal adenocarcinoma epithelial cell line derived from a Dukes?? type C tumor. It harbors a KRAS G13D mutation, a TP53 mutation, and exhibits microsatellite instability, reflecting key genetic features of aggressive colorectal cancers. As an adherent epithelial model, DLD-1 cells are widely employed to study colorectal cancer biology, tumor signaling, and therapeutic responses.
BAG3 encodes a stress-inducible co-chaperone that plays a central role in protein quality control and cell survival. Functioning as a nucleotide exchange factor for Hsp70/Hsc70, BAG3 cooperates with the small heat shock protein HspB8 and the ubiquitin ligase CHIP to mediate chaperone-assisted selective autophagy (CASA). This complex facilitates the recognition and degradation of misfolded proteins and aggregates via the autophagic receptor p62/SQSTM1 and the autophagosomal marker LC3. BAG3 also intersects with apoptosis regulation by inhibiting caspase activation and modulates pro-survival signaling through NF-??B and YAP/TAZ pathways. Its expression is upregulated by diverse stressors, including heat shock, oxidative stress, and inflammatory cytokines such as TNF-??, through transcription factors HSF1 and NF-??B.
In the context of colorectal cancer, BAG3 is frequently overexpressed and contributes to tumor cell survival, apoptosis resistance, and chemoresistance. The DLD-1 cell line, with its mutant KRAS and TP53 background, provides a relevant platform for dissecting how BAG3 coordinates autophagy and survival signaling to sustain malignant phenotypes. Disruption of BAG3 in these cells allows researchers to examine its role in protecting against protein aggregate toxicity, regulating autophagic flux, and maintaining NF-??B and YAP/TAZ transcriptional activity under stress conditions.
Typical experimental applications include assessing autophagic flux using LC3 turnover and p62 degradation assays in the presence of lysosomal inhibitors like bafilomycin A1, quantifying apoptosis by caspase-3/7 activity and annexin V staining, and evaluating drug sensitivity through cell viability assays. The knockout cells are also suitable for co-immunoprecipitation to probe BAG3-containing complexes, immunofluorescence to visualize aggresome formation, and reporter assays for NF-??B and YAP/TAZ activity. This versatile tool supports investigations into autophagy, cancer drug resistance, and stress response signaling. For further technical details, please contact Ascent Research.
