The BCL2L2 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited population derived from the HT29 colorectal adenocarcinoma line, with targeted disruption of the anti-apoptotic BCL2L2 (Bcl-w) gene. This polyclonal knockout product comprises a heterogeneous cell pool generated by direct delivery of editing reagents, avoiding single-cell cloning and thus preserving genetic variability. The resulting loss-of-function model enables functional studies of BCL2L2 within a physiologically relevant epithelial background, offering a robust tool for apoptosis and cancer research.
The host cell line HT29 originates from a human colorectal adenocarcinoma and is characterized by wild-type TP53, microsatellite stability (MSS), and oncogenic mutations in APC and BRAF(V600E). These features render HT29 a widely used model for intestinal epithelial biology and colon cancer, particularly for studying signaling pathways driven by constitutive MAPK activation and chromosomal instability. The epithelial origin supports investigations of cell polarity, adhesion, and tumor progression.
BCL2L2 encodes Bcl-w, an anti-apoptotic protein that binds and inhibits pro-apoptotic BAX and BAK, thereby blocking mitochondrial outer membrane permeabilization and cytochrome c release. This in turn prevents apoptosome assembly involving APAF1 and caspase-9, suppressing effector caspase-3 activation. BCL2L2 expression is driven by transcription factors such as STAT3, CREB, and NF-??B, which act downstream of PI3K/AKT and MAPK/ERK cascades stimulated by EGF and IL-6. Bcl-w also interacts with BH3-only proteins BIM, BID, and BAD, and anti-apoptotic partners MCL-1 and BCL-xL, orchestrating cellular stress responses.
In HT29 cells, BRAF(V600E) drives constitutive ERK signaling, while wild-type p53 preserves an intact DNA damage response. Disruption of BCL2L2 in this background is expected to sensitize cells to intrinsic apoptosis, potentially counteracting chemoresistance mechanisms prevalent in colorectal tumors. This model therefore allows dissection of the interplay between oncogenic MAPK signaling and mitochondrial apoptosis regulation, enabling assessment of Bcl-w dependency in a therapeutically relevant setting.
This polyclonal knockout population supports diverse applications, including chemosensitivity profiling with agents such as 5-fluorouracil and oxaliplatin, validation of Bcl-2 family inhibitors (e.g., navitoclax), and mitochondrial apoptosis signaling studies. Representative assays encompass Annexin V/PI flow cytometry, caspase-3/7 activity, MTT viability, JC-1 mitochondrial potential measurement, western blotting, and co-immunoprecipitation of Bcl-2 interactors. For detailed specifications, contact Ascent Research.
