The HSPA4 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human HSPA4 gene. This loss-of-function model enables investigation of HSPA4-dependent processes in a genetically heterogeneous knockout background. The product is supplied as a polyclonal pool of HT29 cells bearing targeted gene disruption, providing a versatile platform for functional genomics, cancer biology, and stress-response studies without the constraints of monoclonal selection.
The HT29 host cell line, derived from a 44-year-old female patient with colorectal adenocarcinoma, is a well-established model of non-metastatic colorectal cancer. These epithelial cells retain colon-specific features, including the expression of mucins, and are widely employed for dissecting colorectal tumor biology and intestinal epithelial barrier function. The adherent growth and robust in vitro characteristics of HT29 cells make them ideal for stable knockout generation and a broad range of downstream assays.
HSPA4 encodes a stress-inducible chaperone of the HSP70 family integral to protein folding and the cellular heat shock response. Its transcription is activated by HSF1 under conditions of heat stress, oxidative stress, or HSP90 inhibition. HSPA4 interacts with co-chaperones HSP40 and BAG3, and forms complexes with the androgen receptor and AIF. It functions to suppress JNK-mediated apoptosis through modulation of BAX/BCL2 and enhances androgen receptor transcriptional activity by stabilizing the receptor complex, cross-regulating NF-??B signaling.
Within the colorectal cancer model provided by HT29 cells, HSPA4 knockout allows systematic dissection of molecular mechanisms linking chaperone function to tumor cell survival, thermotolerance, and hormone receptor signaling. Loss of HSPA4 is expected to sensitize cells to proteotoxic stress and chemotherapeutic agents by disrupting protein homeostasis and lowering the apoptotic threshold. This model is thus highly relevant for elucidating drug resistance mechanisms and for evaluating the consequences of impaired androgen receptor signaling in colon epithelial cells.
Researchers can utilize the HSPA4 Knockout HT29 Polyclonal Cells in viability assays under heat shock or chemotherapeutics, apoptosis analyses by annexin V/PI flow cytometry, colony formation studies, and drug sensitivity profiling. Molecular validation is accomplished by RT-qPCR and western blot. This polyclonal knockout pool supports investigations into colorectal cancer progression, protein folding fidelity, and apoptosis/survival pathways, and enables high-throughput screens for HSPA4-dependent vulnerabilities. For further technical details and ordering information, please contact our scientific support team.
