DNAJC16 Knockout MES-OV Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the MES-OV mouse embryonic stem cell line. The targeted disruption of DNAJC16 provides a loss-of-function model for studying mitochondrial co-chaperone function and protein homeostasis. As a polyclonal pool, these cells offer a genetically diverse background while uniformly lacking DNAJC16 expression, suitable for population-based assays and functional screens.
The MES-OV host line originates from 129/Sv mouse blastocysts and maintains pluripotent self-renewal and differentiation capacity into all embryonic lineages. These stem cells exhibit stable karyotype and are widely used for gene editing. Given the reliance of pluripotent stem cells on mitochondrial integrity, this system provides a relevant model for examining how DNAJC16-dependent mitochondrial proteostasis influences stem cell maintenance and differentiation.
DNAJC16 encodes a mitochondrial J-protein co-chaperone that partners with mitochondrial Hsp70 (mtHsp70) to stimulate its ATPase activity, driving the translocation of nuclear-encoded preproteins across the inner membrane via the TIM23 translocase complex. It interacts with additional factors such as Tim44, Pam16, and Pam18, and is regulated by stress-responsive transcription factors ATF5 and CHOP under the mitochondrial unfolded protein response (UPRmt). Loss of DNAJC16 disrupts protein import, impairs respiratory chain complex assembly, and activates proteostatic stress signaling.
In the context of pluripotent cells, DNAJC16 knockout illuminates how mitochondrial protein quality control affects self-renewal and lineage commitment. Mitochondrial respiration is critical for maintaining pluripotency; impaired oxidative phosphorylation due to DNAJC16 deficiency may alter cellular metabolism and UPRmt activation, thereby influencing cell fate decisions. This model enables dissection of these mechanisms during early development and in disease contexts.
These cells are suitable for a range of assays: western blot and immunofluorescence for mitochondrial proteins, co-immunoprecipitation with mtHsp70, JC-1 membrane potential measurements, Seahorse respirometry, proteomic analysis of mitochondrial fractions, and RT-qPCR for UPRmt gene expression. Applications include mitochondrial disease modeling, drug screening for mitochondrial dysfunction, and investigation of UPRmt pathways. For further information, contact Ascent Research.
