The C14orf93 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from HAP1 cells, designed to ablate C14orf93 expression. The polyclonal composition avoids single-cell cloning, thereby preserving genetic heterogeneity and minimizing clonal artifacts, while collectively ensuring loss of functional protein. Disruption of the target gene can be confirmed by immunoblotting or RT-qPCR, establishing these cells as a robust foundation for diverse functional investigations.
The HAP1 cell line is a near-haploid adherent cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype??retaining a single copy of most chromosomes??enables unambiguous genotype?Cphenotype correlations following gene disruption, bypassing the complications of diploid complementation. This genetic simplicity makes HAP1 an ideal platform for loss-of-function screens and knockout studies. Additionally, HAP1 cells retain cancer-relevant signaling networks, providing a physiologically relevant context for leukemia research.
The C14orf93 protein is poorly characterized but predicted to localize to mitochondria, where it is thought to contribute to mitochondrial homeostasis. It may function in concert with mitochondrial import receptors, such as the TOM complex, and oxidative phosphorylation complexes, including NADH dehydrogenase and cytochrome c oxidase. Disruption of C14orf93 could compromise protein import or respiratory chain assembly, thereby altering ATP production and redox balance. The protein might act downstream of mitochondrial stress signals or as a structural cofactor, but its precise molecular interactions and regulatory mechanisms remain unknown.
Knockout of C14orf93 in the HAP1 leukemic background offers a powerful system to study mitochondrial contributions to cancer cell fitness. Leukemia cells depend on mitochondrial metabolism for energy and biosynthesis; thus, disruption of mitochondrial homeostasis may impair proliferation or sensitize cells to apoptosis. Combined with HAP1??s haploid genetics, this model enables clear phenotypic readouts using mitochondrial membrane potential (JC-1) assays, oxygen consumption measurements (Seahorse), and Annexin V/PI apoptosis detection. Furthermore, it can be utilized in synthetic lethal screens to identify compounds that target mitochondrial vulnerabilities, linking C14orf93 to leukemia cell survival.
The C14orf93 Knockout HAP1 Polyclonal Cells are suited for functional genomics, mitochondrial biology, and cancer research. Typical assays include Western blotting for knockout validation, RT-qPCR for gene expression analysis, flow cytometry for apoptosis (Annexin V/PI), MTT proliferation assays, mitochondrial membrane potential (JC-1) measurements, Seahorse metabolic flux analysis, and colony formation. Moreover, they facilitate synthetic lethal screening and dissection of mitochondrial stress pathways. These cells support drug target validation and mechanistic studies, providing a platform to investigate C14orf93 function in leukemia and beyond. For further information, contact Ascent Research.
