The NCOA7 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte cell line. These cells feature targeted disruption of the NCOA7 gene, resulting in a loss-of-function model. As a polyclonal population, they provide a heterogeneous pool of NCOA7-deficient cells, avoiding clonal artifacts and enabling robust functional genomics studies. The CRISPR/Cas9-mediated gene knockout ensures effective ablation of NCOA7 protein expression, facilitating dissection of NCOA7-dependent pathways in B cells.
The Raji host cell line, isolated from a Burkitt??s lymphoma patient, is a human B lymphocyte line with lymphoblastoid characteristics. Raji cells maintain key B cell features including surface immunoglobulin expression and participation in antibody production and immune responses. Extensively employed in B cell biology, apoptosis, and virology research, Raji cells offer a physiologically relevant model for investigating nuclear receptor signaling and innate immunity in B lymphocytes. Their rapid growth and responsiveness to growth factors and cytokines make them an ideal system for studying coactivator functions.
NCOA7 is a nuclear receptor coactivator that enhances transcriptional activation by ESR1 and other nuclear receptors, integrating estrogen and interferon signaling pathways. It is activated by estrogen-bound ESR1 and type I interferon/STAT1 signaling, and it forms complexes with SRC-1, IRF3, ATG16L1, and the V-ATPase. NCOA7 transcriptionally upregulates ESR1 target genes such as GREB1 and PGR, and interferon-stimulated genes like MX1 and OAS1. In the endolysosomal compartment, NCOA7 regulates lysosomal genes LAMP1 and CTSB, thereby influencing autophagy and innate immune responses. In B lymphocytes, NCOA7 coordinates hormone and immune signals to control gene expression and cellular homeostasis.
In the context of Raji B lymphocytes, NCOA7 knockout impairs estrogen-driven transcriptional programs that are implicated in B cell lymphoma growth and survival. The loss-of-function model also attenuates interferon-stimulated gene induction, enabling studies of innate immune signaling defects in lymphomagenesis. Given NCOA7??s involvement in endolysosomal function, these knockout cells provide a platform to examine lysosomal dynamics and autophagy in Burkitt??s lymphoma. Researchers can investigate tamoxifen sensitivity and other drug responses, as well as probe crosstalk between estrogen, interferon, and lysosomal pathways in B cells.
Key applications include NCOA7 quantification via Western blotting, RT-qPCR for downstream targets such as GREB1 and MX1, and estrogen-responsive luciferase reporter assays to measure nuclear receptor activity. Flow cytometry with B cell markers confirms lineage identity, while cathepsin B activity and LysoTracker staining assess lysosomal function. Apoptosis and tamoxifen sensitivity assays explore cell death mechanisms. The polyclonal knockout model allows pathway analysis without clonal selection bias. For additional information, please reach out to Ascent Research.
