This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from Raji B lymphocytes, featuring targeted disruption of the MLLT10 gene (encoding AF10). The polyclonal pool is generated via electroporation of ribonucleoprotein complexes, resulting in a heterogeneous knockout model that captures population-level effects of MLLT10 loss of function. The cells are suitable for investigating the transcriptional and epigenetic roles of AF10 in a malignant B-cell background without confounding clonal selection artifacts.
The Raji host cell line originates from a Burkitt lymphoma patient, maintained as an Epstein-Barr virus (EBV)-positive lymphoblastoid line. These cells exhibit mature B-cell phenotypic markers and retain functional machinery for antigen presentation and immunoglobulin production. The lymphomagenic context provides a physiologically relevant system for studying oncogenic transcription factor complexes and epigenetic dysregulation in B-cell malignancies, including diffuse large B-cell lymphoma and Burkitt lymphoma.
MLLT10/AF10 functions as a transcription cofactor and essential scaffold protein within the DOT1L histone methyltransferase complex. Through its interactions with DOT1L, MLL (KMT2A), menin, and histone H3, AF10 facilitates recruitment of DOT1L to chromatin, promoting dimethylation of histone H3 at lysine 79 (H3K79me2) at target loci. This epigenetic modification is critical for sustaining expression of leukemogenic homeobox (HOX) genes such as HOXA9, along with its cofactor MEIS1 and downstream effector FLT3. The AF10-DOT1L axis serves as a central hub in MLL-rearranged leukemogenesis, where chimeric MLL-AF10 fusions aberrantly activate H3K79 methylation. Disruption of MLLT10 is therefore expected to impair DOT1L catalytic activity, reduce H3K79me2 deposition, and downregulate HOX gene transcriptional programs.
In the Raji B-cell lymphoma background, MLLT10 knockout offers a powerful tool for dissecting AF10-dependent signaling networks. Loss of AF10 cofactor function abrogates DOT1L-mediated H3K79me2 enrichment at HOXA9 promoters, leading to diminished expression of leukemogenic targets and potentially attenuating proliferation and survival of lymphoma cells. This model enables direct interrogation of epigenetic dependencies in B-cell neoplasms and facilitates comparative studies between lymphoid and myeloid leukemogenic mechanisms. It also provides a platform for evaluating DOT1L inhibitors (such as pinometostat) in a B-lymphoma context, bridging acute leukemia research with lymphoid malignancy models.
Key applications include quantitative RT-qPCR quantification of HOXA9 and MEIS1 transcript levels to assess knockdown efficiency, western blot detection of global and locus-specific H3K79me2 changes, flow cytometric profiling of B-cell surface markers (CD19, CD20) and proliferation indicators, cell viability and colony formation assays for functional endpoint analysis, RNA sequencing for global transcriptome mapping, and chromatin immunoprecipitation?CqPCR (ChIP-qPCR) to monitor H3K79me2 occupancy at HOX gene regulatory regions. The model supports drug target validation, mechanistic dissection of AF10-DOT1L?Cchromatin complexes, and preclinical assessment of epigenetic therapies. For technical specifications and ordering information, please contact Ascent Research.
