GRPEL2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the GRPEL2 gene in the HAP1 cell line. This heterogeneous pool of gene-disrupted cells provides a loss-of-function model without clonal selection, suitable for mitochondrial biology and genetic screening applications. The polyclonal format enables phenotyping under near-haploid conditions while avoiding clone-specific artifacts.
The HAP1 cell line is a near-haploid adherent line derived from a male chronic myeloid leukemia (CML) patient. Its single-copy chromosome complement ensures that recessive phenotypes are immediately apparent, making it a clean genetic platform for knockout studies. HAP1 is widely used in CRISPR-based functional genomics, and its CML origin offers a cancer-relevant background for investigating oncogenic signaling, metabolism, and drug responses. The line??s stable karyotype and adherent growth support reproducible imaging and biochemical analyses.
GRPEL2 encodes a mitochondrial nucleotide exchange factor essential for protein import and folding. It catalyzes ADP release from mtHsp70 (HSPA9/mortalin), promoting ATP-driven conformational changes required for TIM23-mediated translocation of precursor proteins. GRPEL2 cooperates with TIM23 subunits TIMM23 and TIMM17A, and import receptors TOMM20 and TOMM22 deliver clients. Downstream, the HSP60/HSP10 chaperonin system completes folding. Expression is regulated by PGC-1?? and NRF1 during mitochondrial biogenesis, and by HSF1 under proteotoxic stress, linking GRPEL2 to the mitochondrial UPR. Knockout disrupts mtHsp70 cycling, causing accumulation of unfolded matrix proteins, reduced import of respiratory chain subunits, and loss of proteostasis.
Disruption of GRPEL2 in HAP1 cells allows dissection of mitochondrial dysfunction in a leukemic background. The CML origin helps explore how compromised protein import impacts cancer cell metabolism, proliferation, and apoptosis. Near-haploidy amplifies phenotypic severity, simplifying genetic interaction studies and synthetic lethality screens. This model is suited for probing the mitochondrial UPR, chaperone network function, and bioenergetic adaptation.
Typical readouts include western blotting for mitochondrial proteins, RT-qPCR for UPRmt markers (e.g., HSP60, CLPP), immunofluorescence for morphology, and Seahorse respirometry to measure oxygen consumption. Protein import and co-immunoprecipitation assays monitor translocase activity and mtHsp70 interaction. RNA-seq captures global transcriptional responses. These tools support studies in mitochondrial disease, cancer, and drug discovery. For more details, contact Ascent Research.
