The MISO1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population offering targeted disruption of the MISO1 gene in the HAP1 human cell line. This heterogenous pool provides a robust loss-of-function model for investigating mitochondrial quality control without clonal isolation, reducing artifacts and capturing population-level responses.
HAP1 is an adherent, near-haploid cell line derived from male KBM-7 chronic myeloid leukemia cells. Its near-haploid karyotype ensures that single-gene disruptions yield clear phenotypes, as no second allele masks the knockout effect. Widely used for genetic knockout and haploid screening, HAP1 cells offer ease of culture and retain myeloid characteristics relevant to hematopoietic and cancer studies.
MISO1 encodes a mitochondrial outer membrane scaffolding protein essential for stress-induced mitophagy. Functioning downstream of the PINK1/Parkin pathway, MISO1 is regulated by HIF1A, AMPK, and reactive oxygen species. It recruits autophagic receptors p62/SQSTM1 and LC3 to damaged mitochondria, facilitating ubiquitin-dependent clearance. MISO1 interacts with PINK1, Parkin, BNIP3, NIX, and LC3, linking oxidative stress, mitochondrial unfolded protein response, and mTOR signaling. Disruption of MISO1 impairs mitophagic flux, leading to accumulation of dysfunctional mitochondria, elevated ROS, and compromised homeostasis.
In the near-haploid HAP1 background, MISO1 knockout yields a strong phenotype amenable to high-throughput screening. The polyclonal population model avoids clonal artifacts and simplifies interpretation of signaling events, making it ideal for identifying genetic or pharmacologic modulators of mitophagy. The absence of a second allele allows robust detection of rescue or exacerbation of the knockout phenotype, supporting mechanistic studies and drug discovery.
These cells enable investigations into mitophagy mechanisms, mitochondrial quality control, and oxidative stress in neurodegeneration and cancer. Compatible assays include mitophagy flux measurement by LC3 turnover, ROS detection, mitochondrial membrane potential analysis, western blot for mitophagy markers (LC3, p62, PINK1), immunofluorescence co-localization, ATP quantification, and flow cytometry for mitochondrial mass. For further information, please contact Ascent Research.
