The CHFR Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the CHFR gene in the human Raji B lymphocyte line. This loss-of-function model enables investigation of mitotic checkpoint regulation and chromosomal instability without clonal selection bias. The polyclonal format provides a heterogeneous genetic background while eliminating wild-type CHFR expression, facilitating robust functional studies.
The Raji cell line is a suspension-adapted human Burkitt lymphoma B cell line that is Epstein-Barr virus (EBV)-positive. It serves as a well-established model for B lymphocyte biology, including antigen presentation, antibody production, and adaptive immunity. The B cell origin makes this knockout particularly relevant for studying hematologic malignancies and EBV-driven oncogenesis, where CHFR inactivation contributes to disease progression.
CHFR is a mitotic stress-sensing E3 ubiquitin ligase that ubiquitinates PLK1 and Aurora A, marking them for proteasomal degradation. This activity is triggered by microtubule stress or DNA damage via ATM/ATR signals, delaying mitotic entry to prevent premature chromosome condensation. CHFR interacts with HDAC1, 14-3-3??, and E2 conjugating enzymes; downstream, the APC/C complex and cyclin B1 orchestrate mitotic progression.
In Raji cells, CHFR knockout mimics the frequent promoter methylation observed in colorectal, gastric, and lung cancers. Loss of checkpoint control leads to advanced mitotic entry and chromosomal instability, hallmarks of aggressive tumors. This model is invaluable for studying how checkpoint deficiency cooperates with EBV in lymphomagenesis and for evaluating sensitivity to microtubule-targeting agents like paclitaxel and vincristine.
This polyclonal knockout population supports diverse assays: Western blot for CHFR, PLK1, phospho-histone H3; flow cytometry for cell cycle and mitotic index; immunofluorescence of mitotic spindles; co-immunoprecipitation for CHFR-substrate interactions; RT-qPCR for CHFR expression; and drug sensitivity assays with taxanes or vinca alkaloids. These applications help dissect mitotic checkpoint pathways and drug resistance mechanisms. For more information, contact Ascent Research.
