DHX34 Knockout Raji Polyclonal Cells represent a population of Raji B lymphocytes that have been engineered via CRISPR/Cas9-mediated gene disruption to introduce loss-of-function mutations in the DHX34 locus. This polyclonal knockout cell model provides a heterogeneous pool of edited cells, enabling robust functional studies of DHX34-dependent nonsense-mediated mRNA decay (NMD) in a human lymphoma background.
The Raji cell line is an EBV-positive Burkitt lymphoma-derived B-lymphocyte model that is extensively employed in investigations of B-cell biology, lymphomagenesis, and host-virus interactions. Its documented genomic and transcriptomic landscape, coupled with well-characterized signaling pathways, makes Raji cells a suitable host for dissecting RNA metabolism and translational control mechanisms in the context of B-cell malignancies.
DHX34 encodes a DEAH-box RNA helicase that is a core component of the NMD machinery. Upon recognition of a premature termination codon (PTC) by the terminating ribosome, DHX34 assembles into the SURF complex, which includes the SMG1 kinase, the RNA helicase UPF1, and the eukaryotic release factors eRF1 and eRF3. DHX34 catalyzes ATP-dependent unwinding of RNA secondary structures, which enables phosphorylation of UPF1 by SMG1 and recruitment of the SMG5-SMG7 heterodimer. This cascade directs endonucleolytic cleavage and exonucleolytic decay of aberrant transcripts encoding truncated or potentially toxic proteins. Downstream targets include NMD substrates such as GADD45B, ATF4, and SRSF2, while regulatory inputs derive from PTC-bearing mRNAs and the exon junction complex.
In Raji B lymphocytes, disruption of DHX34 abrogates a critical node of the NMD pathway, leading to stabilization of PTC-containing transcripts that might otherwise be eliminated. This perturbation can unmask aberrant gene expression programs that contribute to lymphomagenesis, immune evasion, or viral latency. Because Raji cells harbor Epstein-Barr virus, the DHX34 knockout model additionally offers a unique system to investigate how viral factors intersect with host NMD to influence B-cell transformation and immune signaling.
Researchers can employ DHX34 Knockout Raji Polyclonal Cells to interrogate NMD dynamics through a variety of experimental approaches, including western blot analysis of DHX34 abundance and UPF1 phosphorylation status, RT-qPCR quantification of endogenous NMD substrates such as GADD45B and ATF4, and transcriptome-wide RNA-seq profiling to identify novel NMD targets in lymphoma. Co-immunoprecipitation assays enable mapping of DHX34 interactions with UPF1, SMG5, SMG7, and eRF factors, while luciferase-based PTC reporters provide a direct readout of NMD efficiency. Functional studies may include flow cytometric assessment of cell cycle progression or apoptosis following DHX34 loss, proliferation assays, and drug sensitivity screens with translational inhibitors to evaluate NMD-targeted therapeutic strategies. The polyclonal nature of the knockout population captures the cellular heterogeneity inherent in cancer models, facilitating robust and reproducible data collection. For technical information or ordering details, please contact Ascent Research.
