The DNAJC5 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered for loss-of-function analysis of the DNAJC5 gene. This product comprises a heterogeneous pool of K-562 cells carrying targeted gene disruption, providing a powerful model to study DNAJC5-dependent mechanisms without the biases associated with clonal selection. The polyclonal nature ensures representation of multiple editing outcomes, enabling robust assessment of gene function in a mixed cellular context relevant to heterogeneous cancer cell populations.
The host K-562 line is a suspension lymphoblastoid cell line originally derived from a 53-year-old female with chronic myelogenous leukemia in blast crisis. K-562 cells exhibit a hematopoietic progenitor-like phenotype, expressing markers of both erythroid and myeloid lineages, and are widely used as a model system for leukemia biology and cytotoxicity assays. Their rapid growth in suspension and well-characterized signaling pathways make them a convenient and reproducible platform for gene editing and subsequent phenotypic screening in hematopoietic research.
DNAJC5 encodes cysteine string protein (CSP), a co-chaperone essential for synaptic vesicle exocytosis and protein homeostasis. CSP is regulated by heat shock factor 1 (HSF1) and cellular stress signals, and it directly interacts with Hsc70/HSPA8 and SGTA to prevent protein aggregation. Functionally, CSP chaperones SNARE proteins such as SNAP-25, syntaxin-1, and VAMP2, thereby modulating SNARE-mediated vesicle fusion. Disruption of DNAJC5 impairs this chaperoning activity, altering SNARE complex assembly and stress response pathways, which can lead to dysregulated exocytosis and proteostasis.
In the K-562 hematopoietic context, this polyclonal knockout model enables investigation of CSP functions beyond neuronal tissues, where it is most studied. The loss of CSP can reveal roles in SNARE-dependent processes that may affect leukemia cell signaling, survival, or secretory activity. Moreover, because DNAJC5 mutations are linked to neuronal ceroid lipofuscinosis type 4 (CLN4), a neurodegenerative disorder, the K-562 knockout population provides a tractable non-neuronal system to explore underlying molecular pathologies, including protein aggregation and chaperone network dysfunction, within a genetic background amenable to high-throughput manipulation.
This polyclonal knockout cell product supports a broad range of applications, including dissection of chaperone-mediated protein homeostasis, elucidation of DNAJC5-dependent stress responses, and drug screening for CLN4-related neurodegeneration. Researchers can employ western blot and RT-qPCR to confirm CSP disruption, co-immunoprecipitation to assess Hsc70 interactions, and heat shock response assays to evaluate stress pathway alterations. Additionally, flow cytometry for vesicle release markers and viability assays enable functional readouts in cytotoxicity studies. For ordering and technical inquiries, please contact Ascent Research.
