The CLPX Knockout Raji Polyclonal Cells consist of a polyclonal Raji B lymphocyte population with CRISPR/Cas9-mediated disruption of the CLPX gene. This loss-of-function model enables investigation of the mitochondrial ATP-dependent protease CLPX in a human B-cell context, without clonal selection, providing a versatile system for functional studies.
Raji is an EBV-positive lymphoblastoid cell line derived from Burkitt lymphoma, widely used to study B-cell malignancies. As a suspension cell line, Raji facilitates high-throughput handling and is compatible with large-scale genetic screens, making it an ideal host for gene-editing studies focused on mitochondrial biology and proteostasis in lymphoma.
CLPX encodes the regulatory subunit of the ClpXP mitochondrial protease complex. It assembles with the peptidase ClpP to form an ATP-dependent proteolytic machine that degrades misfolded matrix proteins and processes the replicative helicase Twinkle, linking protein quality control to mtDNA replication. Transcription of CLPX is upregulated by the stress-responsive factors ATF5 and CHOP in response to reactive oxygen species and unfolded protein accumulation. The ClpXP complex interacts with the mitochondrial chaperones Hsp70 and Hsp60, which facilitate substrate delivery, and its activity is embedded in a network that includes the UPRmt regulators SIRT3 and FOXO3, as well as mtDNA maintenance components TFAM and POLG.
In Raji cells, CLPX disruption impairs mitochondrial proteostasis and mtDNA maintenance, sensitizing cells to mitochondrial stress and apoptosis. The EBV-positive background adds relevance for studying how viral proteins influence mitochondrial pathways. This knockout model is suited for dissecting mitochondrial quality control in B-cell lymphoma, exploring synthetic lethal interactions, and identifying vulnerabilities in EBV-driven lymphomagenesis. For example, loss of CLPX may alter sensitivity to mitochondrial-targeted drugs, and the model can be used to screen for factors essential only in the absence of functional ClpXP.
Representative experimental workflows include quantitative PCR for mtDNA copy number to assess mitochondrial genome stability; Western blotting of oxidative phosphorylation (OXPHOS) complexes to evaluate respiratory chain integrity; Annexin V/propidium iodide staining to quantify apoptosis following treatment with the mitochondrial uncoupler CCCP; enzymatic assays for ClpXP proteolytic activity; RNA-sequencing to detect UPRmt gene expression signatures; and flow cytometry using TMRM or JC-1 dyes to measure mitochondrial membrane potential. This polyclonal knockout population provides a flexible resource for both targeted mechanistic studies and unbiased screening approaches in mitochondrial biology and B-cell cancer. For further information, please contact Ascent Research.
