BAD Knouckout DLD-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the DLD-1 human colorectal adenocarcinoma cell line. These cells harbor a targeted disruption of the BAD gene, a BH3-only pro-apoptotic member of the BCL2 family. The polyclonal format provides a heterogeneous knockout pool suitable for studying loss-of-function effects without clonal bias.
DLD-1 is a well-characterized epithelial cell line established from a Duke’s type C colorectal adenocarcinoma. It is widely used as a model for colorectal cancer, featuring key mutations in genes such as APC, KRAS, and TP53, which contribute to its tumorigenic properties. DLD-1 cells retain adherent morphology and are responsive to apoptotic stimuli, making them ideal for studying intrinsic apoptotic pathways and chemotherapeutic sensitivity.
BAD is a central pro-apoptotic sensor integrating survival and death signals. Unphosphorylated BAD translocates to mitochondria, binds and inhibits anti-apoptotic BCL2 and BCL-xL, promoting BAX/BAK oligomerization, cytochrome c release, and caspase-9/-3 activation. Survival kinases AKT1, PKA, and ERK phosphorylate BAD at Ser112 and Ser136, creating 14-3-3 (YWHAZ, YWHAH) binding sites that sequester BAD in the cytoplasm. This phosphorylation is modulated by upstream factors IGF-1, IL-3, and death receptors. BAD also interacts with glucokinase and is regulated by PP2A and PIM kinases.
In DLD-1 colorectal cancer cells, BAD knockout removes a critical apoptotic checkpoint, likely conferring resistance to intrinsic apoptosis and enhancing survival under stress. This model enables dissection of apoptosis evasion through PI3K/AKT and MAPK/ERK pathways, and exploration of metabolic functions such as glucose metabolism via glucokinase binding. The polyclonal population provides a heterogeneous background for evaluating therapeutics targeting BCL2 family or upstream kinases.
Researchers can utilize this model in assays including Western blotting for phospho-BAD (pSer112, pSer136), Annexin V/PI apoptosis analysis, JC-1 mitochondrial membrane potential measurement, and cytochrome c release assays. Co-immunoprecipitation can assess BAD??s interactions with 14-3-3 or BCL-xL, and viability assays with 5-FU or oxaliplatin evaluate chemosensitivity. The model supports investigation of targeted therapies, apoptosis resistance, and metabolic reprogramming in colorectal cancer. For further information, please contact Ascent Research.
