The LACTB2 Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the human LACTB2 gene. This product provides a heterogeneous pool of HAP1 cells carrying targeted disruption of LACTB2, enabling researchers to interrogate the functional consequences of LACTB2 deficiency without clonal selection artifacts. The knockout is generated using CRISPR/Cas9-mediated gene disruption, resulting in a robust model for investigating LACTB2-dependent mitochondrial processes.
The host cell line, HAP1, is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) model. HAP1 cells retain the BCR-ABL1 fusion oncogene characteristic of CML and possess a predominantly haploid karyotype, which simplifies genetic manipulation and phenotypic analysis. This unique genetic background makes HAP1 cells a powerful platform for functional genomics, drug screening, and pathway dissection, particularly for genes involved in fundamental cellular processes such as mitochondrial biology.
LACTB2 encodes a mitochondrial endoribonuclease essential for processing precursor mitochondrial transcripts. As a key component of the mitochondrial RNA processing machinery, LACTB2 cleaves tRNA-mRNA junctions, facilitating the maturation of mRNAs and tRNAs required for mitochondrial translation. LACTB2 functions downstream of mitochondrial biogenesis regulators such as PGC-1?? and NRF1, and acts upstream of critical mitochondrial-encoded proteins including MT-CO1 and MT-ND1, which are subunits of respiratory chain complexes. It interacts with several RNA-processing and ribosomal factors, including MRPL58, GRSF1, DHX30, and PNPT1, forming a network that coordinates mitochondrial gene expression. Knockout of LACTB2 leads to accumulation of unprocessed transcripts and severe impairment of mitochondrial protein synthesis, ultimately causing respiratory chain deficiency and energy failure.
In the HAP1 cellular context, disruption of LACTB2 provides a physiologically relevant model of mitochondrial translation deficiency. The near-haploid nature of HAP1 cells allows for unambiguous genotype-phenotype correlations, as the effect of LACTB2 loss is not masked by a second allele. This knockout model recapitulates features of combined oxidative phosphorylation deficiency and multisystem mitochondrial diseases, offering a simplified system to dissect the molecular pathology of mitochondrial disorders. Furthermore, the BCR-ABL1-driven CML background permits investigation into intersections between oncogenic signaling and mitochondrial metabolism.
Researchers can employ this polyclonal knockout population in diverse experimental designs. Typical applications include RT-qPCR analysis of mitochondrial transcript processing, Western blotting to assess mitochondrial-encoded proteins such as MT-CO1, Seahorse respirometry to quantify oxidative phosphorylation, and fluorescence microscopy to evaluate mitochondrial morphology. Additional assays like RNA immunoprecipitation and Blue Native PAGE for complex assembly further expand utility. These cells support functional genomics, drug screening for mitochondrial vulnerabilities, and cancer metabolism studies. For further information and technical support, please contact Ascent Research.
