HSPA1L Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population featuring disruption of the HSPA1L gene in the HT29 colorectal adenocarcinoma epithelial cell line. This knockout model provides a genetically heterogeneous pool for studying loss of HSPA1L function, maintaining diverse editing outcomes without clonal selection bias. As a polyclonal preparation, it is suitable for pooled functional analyses and screening applications requiring representation of multiple genotypes. The CRISPR/Cas9-mediated gene knockout abolishes HSPA1L protein expression, enabling dissection of its roles in stress responses and protein homeostasis.
The HT29 cell line, isolated from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female, serves as a well-established model for colorectal cancer and intestinal epithelial biology. HT29 cells retain undifferentiated epithelial characteristics under standard conditions but can differentiate into enterocyte-like cells with polarized barrier function. This plasticity permits investigation of epithelial differentiation, tumorigenesis, and drug transport, making the knockout system valuable for colorectal cancer research and pharmaceutical testing.
HSPA1L encodes a stress-inducible HSP70 family chaperone transcriptionally controlled by HSF1 and activated by cellular stressors, including heat, oxidative, and ER stress (IRE1, PERK, ATF6). It collaborates with co-chaperones such as DNAJ (HSP40), BAG family members (BAG3, BAG4), HOP/STIP1, and the CHIP/STUB1 ubiquitin ligase to promote ATP-dependent protein folding, prevent aggregation, and direct misfolded clients to the ubiquitin-proteasome or autophagy-lysosome pathways. Downstream, HSPA1L influences NF-??B and JNK signaling, stabilizes p53, and contributes to antigen presentation and immune regulation. These interactions place HSPA1L at the intersection of proteostasis, stress signaling, and cell fate decisions.
In colorectal cancer, HSPA1L knockout in HT29 cells impairs stress resistance and disrupts proteome maintenance, sensitizing cells to chemotherapeutic agents such as oxaliplatin and 5-fluorouracil via altered p53 and JNK pathway activity. Loss of HSPA1L also reduces HSP70-dependent antigen presentation, potentially affecting immune surveillance. The model thus enables mechanistic studies of chaperone-mediated drug resistance, epithelial barrier dysfunction, and immune modulation within a colorectal adenocarcinoma background.
Key applications include Western blotting for HSPA1L and phospho-HSF1, viability assays under proteotoxic stress or heat shock, apoptosis assays (caspase-3/7, Annexin V), and co-immunoprecipitation of chaperone-client interactions. Researchers utilize this knockout model to screen for proteotoxic stress sensitizers, investigate epithelial differentiation defects, and evaluate HSPA1L??s role in immune-chaperone crosstalk. For additional product details or technical support, please contact Ascent Research.
