The PARP14 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the PARP14 gene in human Raji B lymphocytes. This loss-of-function model eliminates PARP14 expression, enabling investigation of its biological roles without altering the native genetic background of the host cell line through clonal selection. The polyclonal format preserves heterogeneous editing events across the population, reflecting a more physiologically relevant knockout setting for pooled functional screens and bulk assays.
The host cell line, Raji, is an EBV-positive Burkitt lymphoma-derived B lymphoblastoid cell line, isolated from a pediatric patient. These cells express surface IgM but lack surface IgG, and they serve as a widely utilized model for studying B cell malignancies, Epstein-Barr virus biology, and B cell receptor signaling. The Raji line retains key features of transformed B cells, making it an appropriate context for examining oncogenic mechanisms and therapeutic vulnerabilities in lymphomas.
PARP14 encodes a mono-ADP-ribosyltransferase that functions as a transcriptional co-activator for STAT6. In the IL-4/STAT6 signaling cascade, ligand binding to IL-4R activates JAK1 and JAK3, leading to STAT6 phosphorylation and nuclear translocation. PARP14 interacts with STAT6 and other co-regulators, including PARP9, DTX3L, HDAC1, and HDAC2, to facilitate the expression of downstream targets such as BCL2L1, MCL1, CCND2, SOCS1, and GATA3. Through its catalytic activity and complex formation, PARP14 promotes a pro-survival transcriptional program, counteracting apoptosis and supporting cell cycle progression. Additionally, PARP14 associates with stress granule components like G3BP1, implicating it in stress granule dynamics and cellular stress responses.
In the Raji B-cell context, PARP14-mediated transcription is critical for IL-4-induced survival and proliferation. Knockout of PARP14 abrogates the STAT6-dependent expression of anti-apoptotic BCL2L1 and MCL1, thereby sensitizing B cells to apoptosis and impairing lymphomagenic potential. This model allows dissection of PARP14??s role in B cell receptor and NF-??B pathway crosstalk, and in the maintenance of B cell malignancies. Furthermore, as PARP14 is implicated in immune regulation and inflammatory disorders, the Raji knockout cells provide a tractable system to explore how ADP-ribosylation influences oncogenic signaling and immune evasion in EBV-positive lymphomas.
Researchers can employ these polyclonal knockout cells for a spectrum of applications: validating PARP14 as a drug target in B cell lymphomas, screening small-molecule inhibitors of PARP14 catalytic activity or protein interactions, and probing IL-4/STAT6 signaling dynamics using phospho-flow cytometry, ChIP-qPCR, or co-immunoprecipitation. Functional outcomes can be assessed through proliferation assays (MTT, CellTiter-Glo), apoptosis assays (Annexin V/PI), and transcriptomic analysis via RNA-seq. The cells are compatible with standard biochemical techniques, including Western blotting and RT-qPCR, for confirming target disruption and measuring downstream gene expression changes. For further technical information or to obtain this product, please contact Ascent Research.
