The PCLAF Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B-lymphocyte suspension line. This product offers a genetically heterogeneous pool in which the PCLAF gene (KIAA0101) has been disrupted, enabling loss-of-function studies without clonal isolation. The polyclonal format is ideal for pooled functional genomics, biochemical assays, and experiments where population-level knockout diversity is advantageous. It provides a ready-to-use tool for interrogating DNA damage tolerance and proliferation pathways in a lymphoma-relevant context.
Raji cells were established from a Burkitt’s lymphoma patient and represent an aggressive B-cell malignancy characterized by MYC deregulation. As suspension-adapted lymphoblastoid cells, they retain active DNA replication and repair machinery, making them a pertinent model for studying genome maintenance under oncogenic stress. This host background is particularly suited to examine how MYC-driven replication stress converges on DNA damage tolerance factors such as PCLAF.
PCLAF encodes the PCNA-associated factor, which directly interacts with proliferating cell nuclear antigen (PCNA) and promotes its monoubiquitination at lysine 164 by the E3 ligase RAD18. This modification serves as a molecular switch that recruits translesion synthesis (TLS) polymerases, notably POLH, to stalled replication forks, enabling lesion bypass and DNA damage tolerance. PCLAF is transcriptionally regulated by E2F transcription factors and MYC, thereby coupling cell cycle entry to genome stability pathways. Key interacting partners include PCNA, RAD18, ubiquitin, and POLH, while REV1 and FANCD2 participate in downstream repair events. Disruption of PCLAF thus compromises PCNA ubiquitination and TLS polymerase loading, sensitizing cells to replication-blocking lesions.
Within the Raji lymphoma background, PCLAF knockout uniquely addresses the dependency of MYC-overexpressing cells on damage tolerance for survival. Impaired PCNA ubiquitination and reduced TLS capacity are expected to increase sensitivity to crosslinking agents such as cisplatin and UV mimetics. This model facilitates identification of synthetic lethal relationships and validation of PCLAF as a potential therapeutic target in Burkitt’s lymphoma and related B-cell malignancies, while also revealing how proliferative signals sustain genome integrity under replicative stress.
Typical experimental applications include quantitative Western blot analysis of PCLAF levels and PCNA ubiquitination status, immunofluorescence detection of ??H2AX foci to assess DNA damage, and flow cytometry for cell cycle distribution. Proliferation can be measured via BrdU incorporation, while the comet assay detects DNA strand break accumulation. Drug sensitivity profiling with DNA-damaging agents tests chemosensitization phenotypes. These assays support target engagement studies, DNA repair inhibitor screening, and mechanistic dissection of translesion synthesis in lymphoma. For additional information or custom requests, please contact Ascent Research.
