The MBNL3 Knockout Raji Polyclonal Cells are a heterogeneous Raji B lymphocyte population with CRISPR/Cas9-mediated disruption of the MBNL3 gene. This polyclonal knockout pool models loss-of-function of the muscleblind-like 3 RNA-binding protein in B-cell lymphoma. The cells retain editing diversity, mimicking tumor heterogeneity, and are ideal for functional genomics where clonal variation is undesirable.
Raji cells, an EBV-positive immortalized B lymphocyte line from Burkitt lymphoma, grow in suspension and are widely used for B-cell biology and lymphoma studies. They exhibit constitutive NF-??B signaling and express B-cell markers, providing a relevant system to investigate oncogenic mechanisms. The MBNL3 knockout in this background enables study of aberrant RNA processing in lymphomagenesis.
MBNL3 is a post-transcriptional regulator that modulates alternative splicing, polyadenylation, and mRNA stability by binding YGCY motifs in target transcripts. Its expression is induced by the oncogenic transcription factors MYC and NF-??B, and it is further regulated by cellular stress signals. MBNL3 forms complexes with spliceosomal components and splicing factors such as U2AF2, SRSF1, and HNRNPA1, and functionally interacts with its paralog MBNL1. It directly controls the alternative splicing and expression of key apoptosis regulators including BCL2L11 (Bim), MCL1, and PTEN, along with the cell cycle kinase CDK2. Disruption of MBNL3 thus leads to an imbalance in pro- and anti-apoptotic BCL2 family members, altered NF-??B p65 transcriptional activity, and modulation of caspase-3 cleavage, linking RNA processing to B-cell survival pathways.
Within the Raji lymphoma background, MBNL3 knockout directly alters the splicing of BCL2L11, producing a shift in isoform ratios that sensitizes cells to apoptotic cues. Concurrent dysregulation of MCL1 and PTEN further dampens NF-??B pathway output, undermining the constitutive survival signaling that Raji cells depend on for proliferation and EBV latency maintenance. This polyclonal knockout thus captures how splicing factor loss impacts the oncogenic network in B-cell lymphoma, offering a system to dissect RNA processing contributions to tumor maintenance.
The MBNL3 Knockout Raji Polyclonal Cells are suitable for detailed functional studies. Researchers can employ RNA sequencing and splicing-sensitive PCR to map MBNL3-dependent splice isoforms, and Western blotting to measure changes in BCL2L11, CDK2, PTEN, and other targets. Flow cytometry with Annexin V and MTT assays provide quantitative readouts of apoptosis and proliferation. NF-??B reporter assays clarify the crosstalk between MBNL3-mediated RNA processing and transcription factor activity, while RNA immunoprecipitation (RIP) enables identification of MBNL3-bound transcripts. The polyclonal nature of the pool also supports small-molecule splicing modulator screens and investigations into drug resistance mechanisms. Please contact Ascent Research for further information.
