DNAJC7 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that disrupts DNAJC7 gene expression. This loss-of-function model eliminates the J-domain co-chaperone, enabling study of HSP70/HSP90-mediated protein folding. The polyclonal format provides a heterogeneous pool of edited alleles for robust functional studies without single-cell cloning artifacts.
The parental HAP1 cell line is a near-haploid human hematopoietic model derived from the KBM-7 chronic myeloid leukemia line. HAP1 cells grow in suspension and have a single chromosome set, which simplifies gene editing outcomes and ensures unambiguous phenotypes. This haploid background is widely used for high-throughput genetic screens and drug sensitivity profiling.
DNAJC7 acts as a co-chaperone partnering with HSPA8 and HSP90AA1 to fold steroid hormone receptors and kinases. It is activated by HSF1 under stress and functions downstream of steroid signals to mature the glucocorticoid receptor. DNAJC7 interacts with HSP70/HSP90 complexes and regulates client protein stability; its disruption impairs the unfolded protein response and chaperone-mediated autophagy. Knockout cells exhibit reduced hormone-dependent transcription and compromised kinase maturation, offering a platform to dissect chaperone signaling.
In HAP1 cells, DNAJC7 knockout creates a fully penetrant loss-of-function model for studying proteostasis failure in a leukemic background. The absence of a second allele eliminates residual co-chaperone activity, exposing dependencies on the HSP70/HSP90 system. This setting is well-suited for testing responses to proteasome inhibitors and HSP90 antagonists, and for identifying synthetic lethal interactions in cancer cells with compromised chaperone networks.
Key applications include western blotting for client protein degradation (e.g., glucocorticoid receptor, kinases), luciferase reporter assays for steroid receptor activity, and proteotoxicity assays. RNA-seq reveals transcriptional stress responses, and proliferation assays assess fitness under proteotoxic stress. This model facilitates synthetic lethal screens to identify vulnerabilities arising from chaperone disruption. For further details, contact Ascent Research.
