The C18orf32 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional interrogation of the human C18orf32 gene. This product comprises a heterogeneous pool of HAP1 cells harboring targeted disruptions at the C18orf32 locus, introduced via CRISPR/Cas9-mediated gene disruption. The resulting polyclonal knockout model serves as a loss-of-function tool for investigating the biological role of C18orf32, a gene encoding an uncharacterized protein with predicted transmembrane domains. By ablating gene expression across a population, this system enables robust phenotypic analysis without the clonal selection biases inherent to single-cell-derived lines.
The host cell line, HAP1, is a near-haploid human cell line derived from the chronic myeloid leukemia (CML) cell line KBM-7. HAP1 cells retain a hematopoietic progenitor phenotype and carry a predominantly haploid karyotype, which simplifies genetic manipulation and facilitates the generation of unequivocal knockout genotypes. Their origin from a male CML patient provides a relevant background for studies of hematological malignancies, although HAP1 cells have been widely adopted as a versatile platform for functional genomics across diverse biological contexts. The haploid nature of HAP1 cells is particularly advantageous for genetic screens, as it eliminates the need for biallelic targeting and reduces genetic redundancy, thereby increasing the penetrance of loss-of-function phenotypes.
C18orf32 is an orphan gene that encodes a protein of unknown function, distinguished only by in silico predictions of transmembrane domains. Currently, no upstream regulators, downstream effectors, or interacting partners have been identified for this protein, and its contribution to cellular signaling networks remains completely uncharacterized. The absence of functional annotation places C18orf32 among the many human genes awaiting systematic de-orphanization. This knockout cell population provides a clean background to examine potential roles in cellular processes such as proliferation, apoptosis, or mitochondrial homeostasis, as well as to discover molecular interactions through complementation or proteomics approaches.
In the HAP1 haploid background, disruption of C18orf32 offers a unique opportunity to unmask phenotypes that might be masked by a second allele in diploid cells. The near-haploid karyotype ensures that even partial disruption can lead to a complete loss of function at the protein level, enhancing the sensitivity of downstream assays. Researchers can exploit this model to perform unbiased genetic screens??including drug sensitivity, metabolic, or morphological screens??to link C18orf32 to specific biological pathways. Moreover, the hematopoietic origin of HAP1 cells provides a physiologically relevant context for exploring any putative role of C18orf32 in leukemogenesis or hematopoiesis, should such a connection emerge.
Typical research applications for this product include functional characterization of uncharacterized genes, CRISPR-based functional genomics, and haploid genetic screens. The polyclonal knockout cells are well-suited for a range of downstream assays, including CRISPR validation by next-generation sequencing or TIDE analysis, Western blotting to confirm loss of C18orf32 protein (if antibodies become available), RT-qPCR to quantify transcript ablation, and phenotypic assays measuring cell proliferation, apoptosis, or mitochondrial function. By offering a ready-to-use, heterogeneous knockout population, this product accelerates discovery for laboratories investigating novel gene functions. For further details, including customization options, please contact Ascent Research.
