The C9orf85 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional interrogation of the C9orf85 gene. This product is generated by CRISPR/Cas9-mediated gene disruption in the HAP1 human near-haploid cell line, yielding a heterogeneous pool of cells carrying loss-of-function mutations in the target locus. The polyclonal format provides a robust and efficient model system for studying gene function without the need for single-cell cloning, facilitating rapid phenotype screening in a physiologically relevant cellular context.
The host cell line, HAP1, is a human fibroblastoid cell line derived from the male chronic myeloid leukemia KBM-7 line. HAP1 cells exhibit a near-haploid karyotype, which greatly simplifies gene editing by requiring disruption of only a single allele. This characteristic, combined with their adherent growth and stable genome, makes them an ideal platform for generating knockout models, particularly for genes with unknown or poorly characterized functions. The haploid nature reduces genetic complexity and enhances the efficiency of loss-of-function studies.
C9orf85 encodes a protein of currently unknown function; however, domain analyses suggest a potential association with ciliary structures. The protein may be involved in cilium assembly or intraflagellar transport, as it shares features with axonemal and transport components. While its upstream regulators, downstream targets, and interacting partners remain uncharacterized, it is hypothesized to function within the ciliary proteome, possibly interacting with intraflagellar transport proteins or ciliary axonemal proteins. Elucidating its role is critical for understanding ciliary biology and potential ciliopathies.
The HAP1 knockout model provides a powerful system to dissect the function of C9orf85 in a clean genetic background. The haploid genome ensures efficient CRISPR/Cas9-mediated disruption, resulting in a pool of cells with targeted gene inactivation. This model allows researchers to bypass the challenges associated with diploid editing and enables direct assessment of phenotypic consequences. Given the gene??s possible involvement in cilium structure, the HAP1 knockout cells are particularly suited for high-content screening of ciliary phenotypes, motility assays, and transcriptomic profiling to uncover its mechanistic role.
Typical applications include ciliogenesis assays to evaluate cilium formation and maintenance, immunocytochemistry using antibodies against ciliary markers such as acetylated tubulin and Arl13b, and cell motility assays to assess flagellar or ciliary function. Additionally, RNA sequencing can be performed to identify transcriptional changes associated with C9orf85 loss. These polyclonal knockout cells are valuable for genetic screens, functional genomics studies, and exploratory research into the molecular basis of ciliopathies. For further details, technical support, or customization options, please contact Ascent Research.
