OMA1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human Raji B lymphocyte line, carrying targeted disruption of the OMA1 gene. This loss-of-function model provides a robust system for dissecting OMA1-dependent mitochondrial stress signaling in a lymphoid background without the constraints of single-cell clonal selection, ensuring representation of the polyclonal response.
Raji is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma B lymphocyte cell line widely utilized to investigate humoral immunity, antibody production, and antigen presentation. Derived from a pediatric patient, Raji cells retain hallmarks of transformed B cells, including surface immunoglobulin expression and EBV latency, making them a physiologically relevant host for exploring oncogenic signaling, immune cell metabolism, and virus-host interactions.
OMA1 encodes a mitochondrial inner membrane zinc metalloprotease activated by mitochondrial membrane depolarization, reactive oxygen species (ROS), and unfolded protein stress. Upon activation, OMA1 cleaves the dynamin-like GTPase OPA1 at S1 and S2 sites, disrupting long OPA1 isoforms to inhibit fusion and promote mitochondrial fragmentation. Concurrently, OMA1 proteolyzes DELE1, which translocates to the cytosol to engage the heme-regulated inhibitor (HRI/EIF2AK1) kinase. HRI phosphorylates eIF2??, triggering translational reprogramming and selective ATF4 induction to execute the integrated stress response (ISR). OMA1 activity is modulated by interacting proteases PARL and AFG3L2 and cooperates with YME1L in inner membrane quality control, positioning it upstream of effectors DNM1L, MFN1/2, and transcriptional programs regulating apoptosis.
In the Raji B lymphocyte context, OMA1 disruption uncouples mitochondrial dysfunction from cellular adaptation pathways, allowing precise dissection of OPA1-dependent morphology, ISR activation, and apoptotic thresholds. This knockout illuminates how EBV-mediated metabolic rewiring intersects with mitochondrial proteostasis, revealing roles for OMA1 in B cell receptor signaling, antigen presentation fidelity, and lymphoma survival under genotoxic or metabolic stress.
This polyclonal knockout population supports diverse experimental strategies, including Western blotting for OMA1 and OPA1 isoforms, immunofluorescence for mitochondrial morphology via TOM20 staining, flow cytometry using JC-1 or TMRE to gauge membrane potential, and apoptosis assays with Annexin V/PI. Further applications encompass co-immunoprecipitation of DELE1 or PARL, RNA-seq transcriptomics after CCCP or FCCP challenge, and ATF4 reporter assays to monitor ISR engagement. These cells are ideal for drug screening targeting mitochondrial proteostasis, exploring cancer metabolism, and dissecting EBV-host interplay. For further technical information or custom requests, please contact Ascent Research.
