The CRYZ Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B lymphoblastoid line, engineered to disrupt the CRYZ gene encoding quinone oxidoreductase. This polyclonal model provides a heterogeneous loss-of-function system that retains Raji cell characteristics while enabling CRYZ functional studies. The use of a polyclonal population avoids clonal selection artifacts, offering a representative genetic background. CRISPR/Cas9-mediated gene disruption generates a mixed cell pool with collective loss of CRYZ expression, designed for reproducible experiments in oxidative stress and B-cell lymphoma research.
Raji is a human B lymphoblastoid cell line originating from a Burkitt lymphoma patient. It is Epstein-Barr virus (EBV)-positive and displays mature B lymphocyte features, serving as a standard model for B-cell biology, signal transduction, and lymphoid malignancies. Raji cells proliferate rapidly in suspension, with a well-characterized genome, facilitating high-throughput screening and mechanistic studies of apoptosis and transformation. The lymphoblastoid context also supports EBV-oncogenesis research, providing a physiologically relevant host to investigate CRYZ in B-cell oxidative defense.
CRYZ (crystallin zeta) is a cytosolic NADPH-dependent quinone oxidoreductase that catalyzes two-electron reduction of quinones to hydroquinones, protecting against oxidative stress. It is transcriptionally regulated by NRF2 (NFE2L2), which, upon release from KEAP1 under stress, binds the antioxidant response element in the CRYZ promoter. CRYZ acts downstream of NRF2 and upstream of reduced quinones, diminishing ROS production. It shares functional overlap with phase II enzymes like NQO1, together detoxifying reactive electrophiles. NADPH is the essential cofactor.
Knockout of CRYZ in Raji cells provides a model to study quinone detoxification in B-cell homeostasis and lymphoma. Burkitt lymphoma cells experience high oxidative stress; CRYZ loss may increase sensitivity to quinone-based drugs or oxidative insults, revealing chemoresistance. The NRF2-CRYZ axis can be interrogated for its role in redox balance and drug sensitivity. Additionally, with CRYZ linked to autosomal recessive cataract, these cells enable investigation of oxidative stress pathways relevant to cancer and degenerative diseases. The polyclonal format minimizes clonal adaptation artifacts.
Applications include Western blotting to validate CRYZ loss and assess NQO1 compensation; DCFDA assays for ROS measurement; cell viability tests under H2O2 or menadione to gauge oxidative stress resistance; drug sensitivity screens with quinone-based agents; flow cytometry for annexin V apoptosis; and RT-qPCR profiling of NRF2 targets (e.g., NQO1, HMOX1) for transcriptional reprogramming. For further information, please contact Ascent Research.
