The PCIF1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population enabling disruption of the PCIF1 gene in the human Raji B lymphocyte line. This polyclonal pool offers a heterogeneous collection of edited alleles, avoiding clonal selection bias and providing a robust system for functional studies of the cap-specific adenosine N6-methyltransferase PCIF1.
The Raji cell line is derived from a Burkitt lymphoma patient and is Epstein-Barr virus (EBV)-positive. These B lymphocytes maintain key functions including antibody production, antigen presentation, and immune memory. Raji cells are extensively used in immunology and cancer research due to their rapid growth, genetic tractability, and relevance to lymphomagenesis.
PCIF1 encodes a cap-specific adenosine N6-methyltransferase that is recruited to the phosphorylated C-terminal domain (CTD) of RNA Polymerase II during transcription. This enzyme catalyzes N6-methyladenosine (m6A) modification at the first nucleotide of the mRNA cap, a mark recognized by eukaryotic translation initiation factor 4E (eIF4E). The cap-adjacent m6A modulates translational efficiency and mRNA stability, impacting gene expression. PCIF1 functions downstream of transcription initiation factors and the cap-binding complex, and its activity influences ribosome recruitment and protein synthesis. The pathway includes RNA Pol II CTD phosphorylation, PCIF1 recruitment, cap m6A methylation, eIF4E recognition, and translational control.
In the Raji B lymphocyte context, PCIF1 knockout provides a model to study epitranscriptomic regulation in Burkitt lymphoma and EBV-associated pathologies. Since Raji cells carry latent EBV, this system allows investigation of viral mRNA cap modification and host translation control in lymphomagenesis. The knockout also enables dissection of PCIF1’s role in B cell gene expression programs related to antibody production and immune memory.
Key applications include studying cap-adjacent m6A epitranscriptomic modifications, translation regulation, and B-cell lymphoma biology. Researchers can employ Western blotting for PCIF1 depletion, RT-qPCR for target mRNAs, anti-m6A RNA immunoprecipitation, cap methylation assays, translation reporter assays, and RNA-seq. Flow cytometry for B-cell surface markers, proliferation, and apoptosis assays can assess phenotypic outcomes. For more information, please contact Ascent Research.
