CKAP4 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Burkitt’s lymphoma B lymphocyte line Raji. This product provides a loss-of-function model for the CKAP4 gene, generated through CRISPR/Cas9-mediated gene disruption. The polyclonal format preserves genetic heterogeneity, offering a versatile tool to study gene function without clonal selection bias. Researchers can employ this model to investigate CKAP4-dependent signaling, ER morphology, and cytoskeletal organization in a well-characterized B cell context, enabling both targeted mechanistic studies and unbiased phenotypic screens.
The Raji cell line is a widely used model in immunology and cancer research, originating from an EBV-positive Burkitt’s lymphoma. Raji cells retain characteristic features of mature B lymphocytes, including expression of surface immunoglobulins and CD markers, and are commonly used to examine immune functions, antibody production, and lymphomagenesis. Their robust proliferation, ease of culture, and susceptibility to genetic manipulation make them an ideal host for gene knockout studies. The EBV-positive background also renders them relevant for investigating viral oncogenesis and host-virus interactions.
CKAP4 encodes an endoplasmic reticulum (ER)-resident type II transmembrane protein that serves as a high-affinity receptor for the secreted glycoprotein DKK1. Upon DKK1 binding, CKAP4 activates the PI3K/AKT signaling axis, leading to phosphorylation of AKT and subsequent activation of downstream effectors such as mTOR, c-Myc, and stabilization of ??-catenin. CKAP4 also directly anchors the ER to microtubules through its cytosolic domain, a function critical for maintaining ER morphology and spatial distribution. At the ER membrane, CKAP4 interacts with GRP78/BiP, Sec61, and other translocon components, and it forms complexes with mitochondrial proteins VDAC1 and IP3R, bridging ER-mitochondrial communication. Upstream, CKAP4 is regulated by ER stress inducers like tunicamycin and thapsigargin, and by IL-6/STAT3 signaling. Thus, CKAP4 integrates extracellular signals, ER stress responses, and cytoskeletal dynamics to control cell proliferation, survival, and migration.
In the Raji B lymphocyte background, CKAP4 knockout provides a unique platform to dissect the role of DKK1/CKAP4/PI3K/AKT signaling in lymphoma biology. Since Raji cells are derived from aggressive lymphoma, loss of CKAP4 may alter proliferative and anti-apoptotic signaling, potentially modifying tumor cell fitness. The polyclonal population mimics the heterogeneity of tumor cell responses and allows assessment of CKAP4??s impact on ER structure, microtubule organization, and secretory pathway function in a B cell context. Additionally, the EBV-positive status of Raji cells enables exploration of potential crosstalk between viral oncoproteins and CKAP4-mediated pathways.
Typical research applications include assessment of CKAP4-dependent DKK1 signaling by Western blot analysis of AKT phosphorylation and its downstream targets mTOR and c-Myc; immunofluorescence staining to examine ER morphology and microtubule networks; and functional assays such as EdU proliferation, MTT viability, Annexin V apoptosis, and migration. Co-immunoprecipitation can verify disrupted CKAP4?CDKK1 or CKAP4?CGRP78 interactions, while RNA-seq reveals transcriptional changes. Flow cytometry allows monitoring of B cell surface markers. This knockout model is also suitable for studying ER stress responses using pharmacological inducers and for high-content screening. For further information, please contact Ascent Research.
