The CDKAL1 Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the CDKAL1 gene. This product provides a heterogeneous pool of Raji B lymphoblastoid cells harboring targeted disruption of CDKAL1 via CRISPR/Cas9-mediated gene editing, enabling researchers to investigate the functional consequences of CDKAL1 ablation in a B-cell context. The polyclonal format offers a cost-effective and rapid route to gene knockdown without the clonal selection bottleneck, suitable for population-level analyses of gene function.
The Raji cell line is an EBV-positive human B lymphoblastoid line originally derived from a Burkitt lymphoma patient, widely used as a model for B lymphocyte biology, immune response, and antibody production. These suspension cells express characteristic B-cell surface markers such as CD19 and CD20 and maintain a stable lymphoid phenotype, making them a robust platform for studying B-cell signaling, lymphomagenesis, and translational control mechanisms. Their EBV-transformed status confers continuous proliferation and ease of culture, facilitating high-throughput genetic screening and drug response assays.
CDKAL1 encodes a tRNA methylthiotransferase that catalyzes the 2-methylthiolation of N6-threonylcarbamoyladenosine (ms2t6A) at position 37 of tRNA-Lys(UUU). This modification is essential for accurate decoding of Lys codons, particularly for proinsulin translation in pancreatic ??-cells. CDKAL1 interacts with iron?sulfur cluster biogenesis machinery and uses tRNA substrates to generate ms2t6A-modified tRNALys, directly impacting proinsulin translation fidelity and insulin processing. Disruption of CDKAL1 causes ribosomal pausing at Lys codons, triggering ER stress marked by CHOP and BiP upregulation, and impairs insulin secretion. Though best characterized in ??-cells, its tRNA-modifying activity likely influences global protein synthesis in B cells, potentially affecting immunoglobulin production and stress responses.
In the Raji B-cell background, loss of CDKAL1 provides a unique opportunity to dissect the interplay between tRNA modification, translational control, and ER stress in lymphoid cells. Given the Raji line??s origin from Burkitt lymphoma, this model allows investigation into how dysregulated protein synthesis contributes to lymphomagenesis and immune dysregulation. The polyclonal knockout population mirrors the genetic heterogeneity of tumor cell pools, making it suitable for studying clonal variation in stress adaptation and for identifying synthetic lethal interactions with compounds that target the ER stress pathway or tRNA modification pathways.
Typical applications include functional studies of CDKAL1 in B cell biology, investigation of tRNA modification effects on protein synthesis and ER stress, and drug screening for tRNA modification inhibitors. Western blotting for CDKAL1 and ER stress markers (CHOP, BiP), RT?qPCR for tRNA modification enzymes, and LC?MS for ms2t6A quantification can be employed. Flow cytometry for B?cell surface markers (CD19, CD20) monitors phenotypic changes, while viability and apoptosis assays evaluate fitness. This B?cell lymphoma model also serves metabolic disorder research, linking tRNA biology to insulin pathways. For further information, contact Ascent Research.
