The GABARAPL1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to provide a loss-of-function model for the GABARAPL1 gene. This product consists of a heterogeneous pool of Raji B lymphocytes harboring targeted disruption of GABARAPL1, enabling robust functional studies without the need for clonal selection. The polyclonal format captures the genetic diversity of edited alleles, making it well-suited for population-level analyses of autophagy and related processes.
The parental Raji cell line is an EBV-positive lymphoblastoid cell line derived from a Nigerian patient with Burkitt’s lymphoma. Widely used in immunology and cancer research, Raji cells exhibit characteristic B-cell surface markers and have been instrumental in studies of B-cell receptor signaling, apoptosis, and viral oncogenesis. Their robust growth and ease of transfection make them a reliable host for gene-editing experiments, particularly in the context of hematological malignancies.
GABARAPL1 is a member of the ubiquitin-like LC3/GABARAP protein family and plays an essential role in selective autophagy. It is conjugated to phosphatidylethanolamine on autophagosomal membranes through a ubiquitination-like cascade involving ATG7 and ATG3, promoting autophagosome closure and subsequent fusion with lysosomes. GABARAPL1 transcription is regulated by the TFEB/TFE3/FOXO3 network and is suppressed by mTORC1 under nutrient-rich conditions. It functions as a key adaptor by interacting with autophagic cargo receptors such as SQSTM1/p62, NBR1, and CALCOCO2/NDP52, thereby facilitating the degradation of damaged mitochondria, protein aggregates, and intracellular pathogens. Its knockout impairs both bulk and selective autophagic flux, leading to accumulation of autophagic substrates.
In Raji B lymphocytes, GABARAPL1 is poised to influence autophagy-dependent survival mechanisms critical for lymphoma cell proliferation and drug resistance. EBV-associated latency programs can modulate autophagy, and the knockout of GABARAPL1 in this background provides a platform to dissect how autophagic flux intersects with oncogenic signaling in B-cell malignancies. The model allows investigation of whether GABARAPL1-mediated selective autophagy contributes to the clearance of oncogenic proteins or damaged organelles, and whether its loss sensitizes cells to chemotherapeutic agents or targeted therapies.
This knockout cell population is suitable for a broad range of autophagy-focused assays. Researchers can assess autophagic flux by monitoring LC3-II and SQSTM1/p62 levels via Western blotting in the presence or absence of lysosomal inhibitors (e.g., chloroquine or bafilomycin A1). Immunofluorescence microscopy can be employed to quantify GFP-LC3 puncta, while co-immunoprecipitation enables mapping of protein?Cprotein interactions within the autophagy network. Functional studies under starvation conditions or ER stress provide insights into metabolic adaptation. The model is applicable to cancer biology, neurodegenerative proteinopathy research, and host?Cpathogen interactions. For further details or customized solutions, please contact Ascent Research.
