The DNAJC5 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells with targeted disruption of the DNAJC5 gene. This polyclonal knockout pool provides a loss-of-function model for studying the co-chaperone CSP??, encoded by DNAJC5. The use of a polyclonal format enables robust gene disruption across the cell population, allowing researchers to perform biochemical and functional assays without clonal isolation. This product is designed for applications requiring population-level knockout effects, such as protein interaction and aggregation studies.
HAP1 is a near-haploid human cell line derived from the male KBM-7 chronic myeloid leukemia line. Its haploid genome makes it particularly suitable for CRISPR-based knockout studies, as disruption of a single allele typically results in complete gene inactivation. HAP1 cells are widely used in functional genomics and have become a standard system for genome-wide screens. Despite their non-neuronal origin, HAP1 cells express conserved components of exocytic and chaperone pathways, enabling the study of genes like DNAJC5 that play critical roles in neuronal functions.
DNAJC5 encodes cysteine string protein ?? (CSP??), a synaptic vesicle co-chaperone that recruits Hsc70 to SNARE proteins including SNAP-25, syntaxin, and VAMP2, as well as synaptotagmin, to facilitate proper folding and exocytosis. CSP?? activity is regulated by heat shock factor 1 (HSF1) and neuronal activity, and it functions upstream of SNARE complex assembly to maintain presynaptic protein homeostasis. Loss of CSP?? leads to SNARE misfolding and aggregation, contributing to synaptic dysfunction. Dominant mutations in DNAJC5 cause adult-onset neuronal ceroid lipofuscinosis (Kufs disease), and CSP?? dysfunction is linked to synucleinopathies, positioning this gene at the intersection of chaperone-mediated protein quality control and neurodegeneration.
In the HAP1 background, knockout of DNAJC5 enables the dissection of CSP??-dependent chaperone mechanisms in a genetically clean and experimentally accessible system. The haploid nature ensures efficient gene disruption, yielding a consistent loss-of-function phenotype suitable for quantitative assays. Although HAP1 cells are not neuronal, they retain core exocytic and chaperone machinery, allowing researchers to study fundamental CSP?? interactions and functions. This model is particularly useful for co-immunoprecipitation of CSP??-Hsc70-SNARE complexes and for monitoring protein aggregation using established biochemical methods, providing a simplified platform to complement studies in neuronal models.
This DNAJC5 polyclonal knockout pool is a powerful tool for neurodegenerative disease research, including modeling Kufs disease and investigating chaperone-mediated synaptic dysfunction. Researchers can employ these cells in western blotting, RT-qPCR, co-immunoprecipitation, immunofluorescence, and protein aggregation assays to elucidate CSP??’s role in SNARE complex maintenance and to screen for modulators of protein misfolding. For further information or to discuss your specific experimental needs, please contact Ascent Research.
