The MBNL2 Knockout Raji Polyclonal Cells product consists of a heterogeneous pool of Raji B lymphocytes that have undergone CRISPR/Cas9-mediated gene disruption at the MBNL2 locus, generating a loss-of-function model for studying muscleblind-like splicing regulator 2. As polyclonal knockout cells, this population contains diverse editing events across the target gene, offering a pooled knockout system suitable for functional genomics and pathway analysis without single-cell cloning.
The host cell line, Raji, is a human Epstein-Barr virus (EBV)-positive lymphoblastoid cell line originally derived from a Burkitt’s lymphoma patient. Raji cells exhibit a mature B cell phenotype, expressing surface immunoglobulins and MHC class II molecules, and are widely used to study B lymphocyte biology, including immune response, antibody production, and antigen presentation. Their rapid growth and susceptibility to transfection make them a practical model for genetic manipulation.
MBNL2 encodes a conserved RNA-binding protein that recognizes YGCY tandem repeat motifs in pre-mRNA, acting as a critical regulator of alternative splicing. It functions within a network including upstream regulators such as MEF2 transcription factor, SRF, and MAP kinase/calcium signaling, and interacting partners like CELF1, RBFOX2, hnRNP H, and MBNL1. Knockout of MBNL2 disrupts the splicing of key downstream targets, most notably TNNT2 and INSR, leading to aberrant expression of their splice isoforms. Additionally, MBNL2 influences immune-related transcripts such as CD45 (PTPRC) and contributes to B cell receptor signaling and mRNA surveillance pathways. Its misregulation is implicated in the RNA toxicity mechanisms underlying myotonic dystrophy types 1 and 2, where sequestration by expanded CUG/CCUG repeats depletes functional MBNL proteins.
In the Raji B cell context, MBNL2 knockout provides a valuable platform to dissect post-transcriptional gene regulation in the immune system. Loss of MBNL2 is predicted to alter the splicing landscape of immunologically relevant genes, potentially affecting B cell receptor signaling strength, proliferation, and differentiation. This model enables the investigation of how RNA-binding protein dysfunction contributes to lymphoid cell abnormalities observed in myotonic dystrophy and other repeat expansion disorders. Moreover, it allows for the study of MBNL2-specific roles independent of MBNL1 and MBNL3, thereby clarifying the unique contributions of each family member.
Typical research applications include transcriptome-wide splicing analysis via RNA-seq, targeted isoform detection by RT-PCR, and validation of protein isoform shifts through western blotting. Flow cytometry can assess surface marker expression changes, while co-immunoprecipitation assays determine altered RNA?Cprotein interaction dynamics. This polyclonal knockout cell model is also suitable for high-throughput drug screening campaigns aimed at identifying small molecules that correct splicing defects in myotonic dystrophy and related disorders. For further information or to inquire about tailored applications, please contact Ascent Research.
