The DENND3 Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of Raji B lymphoblasts with targeted disruption of the DENND3 gene. This knockout model is generated by introducing a Cas9-sgRNA complex to create gene disruption at the DENND3 locus, resulting in a heterogeneous pool of cells lacking functional DENND3 expression. It serves as a versatile tool for studying DENND3-dependent processes in a human B lymphocyte background.
The Raji cell line is a well-established human B lymphoblast line derived from a Burkitt’s lymphoma patient. These cells grow in suspension and are Epstein-Barr virus (EBV)-positive, retaining features of mature B cells such as the ability to produce immunoglobulins. Raji cells are widely used to model B-cell malignancies, adaptive immunity, and antibody production, making them a relevant system for dissecting oncogenic and immune-related pathways.
DENND3 functions as a guanine nucleotide exchange factor (GEF) for the small GTPase Rab12, thereby promoting Rab12 activation and subsequent autophagosome formation. Its activity is tightly regulated by nutrient status and upstream kinases: under autophagy-inducing conditions, ULK1 kinase directly phosphorylates DENND3, enhancing its GEF activity toward Rab12 and facilitating autophagic flux. Consequently, DENND3 acts downstream of the mTORC1/AMPK/ULK1 signaling axis and upstream of Rab12, LC3-II, and p62/SQSTM1, integrating signals that control endosomal trafficking and autophagy. Key interacting partners include Rab12 and ULK1, and the pathway involves ATG13, Beclin-1, and other core autophagy machinery components.
In B-cell lymphomas, autophagy can support survival under metabolic stress, contribute to drug resistance, and modulate immune responses. Given that DENND3 promotes autophagy, its disruption in Raji cells provides a model to investigate how autophagy inhibition affects lymphoma cell viability, antibody secretion, and sensitivity to chemotherapeutics. Moreover, because EBV-positive lymphomas may exploit autophagy for latent infection maintenance, the DENND3 knockout line can shed light on virus?Chost interactions.
Researchers can employ these polyclonal knockout cells to dissect DENND3’s role in autophagy regulation using classic readouts such as LC3 puncta formation, LC3-II turnover in the presence of bafilomycin A1, and p62 degradation. The model is also suited for assessing functional impacts on B-cell receptor signaling, antibody production, and apoptosis via flow cytometry. Furthermore, drug sensitivity profiling and proliferation assays can evaluate the therapeutic potential of targeting DENND3 or downstream autophagy pathways in Burkitt lymphoma and other B-cell malignancies. For additional details and ordering information, please contact Ascent Research.
