The DNAJB2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DNAJB2 gene in the HT29 human colorectal adenocarcinoma cell line. This loss-of-function model enables study of DNAJB2-dependent proteostasis without clonal selection bias, providing a heterogeneous knockout pool for investigating chaperone networks and protein quality control pathways in colorectal cancer.
HT29 cells are a human colorectal adenocarcinoma cell line with epithelial morphology, derived from a 44-year-old female patient. Widely used as a model for intestinal epithelial biology and colorectal cancer, they can differentiate into enterocyte-like cells under specific conditions. This cell line expresses key intestinal markers and is employed in studies of mucosal function, drug absorption, and oncogenic signaling.
DNAJB2 encodes an HSP40 family co-chaperone that partners with HSP70 chaperones, stimulating their ATPase activity to facilitate protein folding, refolding, and degradation. Under stress, its expression is induced by HSF1 and the unfolded protein response sensors ATF6, IRE1, and PERK. DNAJB2 links chaperone machinery to the ubiquitin-proteasome system and autophagy by interacting with ubiquitin, the E3 ligase STUB1/CHIP, and LC3/GABARAP proteins, thereby directing misfolded clients for proteasomal degradation or autophagic clearance.
In HT29 colorectal cancer cells, loss of DNAJB2 impairs management of misfolded proteins, potentially sensitizing cells to proteotoxic stress from hypoxia, nutrient deprivation, or chemotherapy. This enables dissection of proteostasis addiction in tumors and identification of synthetic lethal interactions with other ubiquitin-proteasome or autophagy components. Although DNAJB2 mutations are linked to motor neuron diseases, this epithelial model provides a platform to explore general mechanisms of protein quality control failure relevant to cancer cell survival.
This polyclonal knockout cell population supports Western blot and co-immunoprecipitation studies to assess DNAJB2-HSP70 interactions, proteasome activity assays, and autophagy analysis via LC3/p62 immunofluorescence. This knockout model also permits flow cytometric detection of apoptosis and autophagy markers following treatment with candidate compounds. It facilitates drug screening for proteostasis modulators using cell viability assays under ER or proteasome stress, and RT-qPCR to measure HSF1 target gene expression. For technical inquiries, contact Ascent Research.
