The MGME1 Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited human loss-of-function model targeting the MGME1 gene in a Raji cell background. This product comprises a polyclonal population of Raji cells harboring CRISPR-mediated disruption of MGME1, which encodes a mitochondrial single-stranded DNA exonuclease crucial for mitochondrial DNA (mtDNA) replication and maintenance. The cells are provided as a ready-to-use polyclonal knockout cell pool, suitable for functional genomics, disease modeling, and drug screening applications without the need for clonal isolation.
The Raji host cell line is a human B lymphocyte model derived from a Burkitt’s lymphoma and immortalized by Epstein-Barr virus (EBV). These lymphoblastoid cells grow in suspension and retain key characteristics of the adaptive immune response, including antibody production, antigen presentation, and rapid proliferation. As a well-characterized lymphoid line, Raji cells provide a physiologically relevant system for studying mitochondrial function in the context of immune cell biology, metabolic reprogramming, and malignant transformation.
MGME1 functions as a mitochondrial 5′-flap exonuclease essential for processing DNA intermediates during mtDNA replication. It operates in coordination with the core mitochondrial replisome, including DNA polymerase gamma (POLG), the Twinkle helicase (TWNK), and mitochondrial single-stranded DNA-binding protein (mtSSB). MGME1 activity is regulated by replication stress signals and works downstream of mitochondrial transcription factor A (TFAM). Knockout of MGME1 leads to accumulation of aberrant replication intermediates, reduced mtDNA copy number, and impaired expression of respiratory chain subunits, ultimately compromising oxidative phosphorylation.
In Raji cells, disruption of MGME1 creates a valuable model to examine how mtDNA maintenance defects impact immune cell metabolism and function. Given the high energy demands of proliferating lymphocytes, MGME1 loss-of-function can reveal vulnerabilities in mitochondrial quality control pathways. This polyclonal knockout population is particularly useful for studying mitochondrial DNA depletion and repair in a cancer context, as Raji cells exhibit oncogenic features such as EBV-driven growth. The model enables dissection of the interplay between mtDNA integrity and cellular processes like apoptosis, proliferation, and antigen presentation.
Key applications include mechanistic studies of mtDNA replication disorders such as mitochondrial DNA depletion syndrome 11 (MTDPS11) and neurodevelopmental pathologies. Researchers can employ this model for drug toxicity testing, screening for mtDNA repair modulators, and functional rescue experiments. Typical assays involve mtDNA copy number measurement by qPCR, mitochondrial protein immunoblotting, respirometry (Seahorse), immunofluorescence for mitochondrial markers, flow cytometry for mitochondrial mass, apoptosis profiling, and RNA-seq for mitochondrial gene expression. Long-range PCR can be used to detect mtDNA deletions. For additional technical details or custom requests, please contact Ascent Research.
