The ELAC1 Knockout HAP1 Polyclonal Cells offer a CRISPR/Cas9-mediated polyclonal knockout population targeting ELAC1 within the HAP1 cell line, generating a loss-of-function model for exploring tRNA 3′-processing defects. These cells are supplied as a heterogeneous mixture of edited alleles, providing a robust experimental system that avoids clonal selection biases while enabling population-level functional studies.
HAP1 is a near-haploid chronic myeloid leukemia cell line derived from the male KBM-7 line. As a leukemic progenitor cell model, it is extensively used in haploid genetic screens due to its single chromosomal set, which eliminates the confounding effects of diploid gene redundancy. This genetic simplicity allows for precise gene disruption and unambiguous genotype-phenotype correlation, making HAP1 an optimal host for studying fundamental processes like RNA metabolism and mitochondrial function.
ELAC1 encodes a tRNA 3′-endonuclease that catalyzes the removal of the 3′ trailer from precursor tRNAs, a critical step in generating mature, functional tRNAs. Its expression is under the control of key regulators including MYC, which enhances tRNA synthesis, and E2F1, which links tRNA biogenesis to cell cycle progression. ELAC1 collaborates with the RNase P complex and interacts with RRP1B to process specific substrates such as tRNA-His and tRNA-Ser. Disruption of ELAC1 leads to accumulation of unprocessed pre-tRNAs, depletion of mature tRNA pools, and consequent impairment of both mitochondrial and cytosolic translation. This defect particularly compromises mitochondrial protein synthesis, resulting in oxidative phosphorylation deficiency and recapitulating molecular pathologies seen in combined oxidative phosphorylation deficiency 17 (COXPD17).
In the HAP1 background, ELAC1 knockout provides a powerful model to dissect the cellular consequences of defective tRNA maturation. The near-haploid genome simplifies the interpretation of metabolic phenotypes, directly connecting ELAC1 dysfunction to bioenergetic collapse. This system recapitulates the mitochondrial translation defects characteristic of COXPD17, enabling mechanistic studies of how pre-tRNA accumulation alters mitochondrial respiration, reactive oxygen species production, and cell viability under stress conditions.
These polyclonal knockout cells are suitable for a variety of research applications, including detailed characterization of tRNA processing pathways, high-content screening for pharmacological rescue of tRNA maturation, and haploid genetic modifier screens to identify synthetic lethal interactions. Assays commonly deployed with this model include RT-qPCR for pre-tRNA quantitation, Northern blotting for mature tRNA profiling, mitochondrial oxygen consumption rate measurements, western blotting for mitochondrial-encoded proteins, and proliferation assays. For additional information or ordering inquiries, please contact Ascent Research.
