The ARL3 Knockout Jurkat Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal population of human Jurkat T lymphocytes carrying targeted disruption of the ARL3 gene. This ready-to-use knockout model provides a loss-of-function tool for investigating ARL3-dependent processes in a T-cell context, without the need for clonal isolation. The polyclonal format captures a spectrum of editing outcomes, enabling robust analysis of ARL3 function while mitigating potential clonal artifacts.
Jurkat cells, an immortalized human T-lymphocyte line originally established from a patient with acute T-cell leukemia, are a principal model system for dissecting T-cell receptor signaling, apoptosis, and cytokine responses. Their suspension growth, rapid proliferation, and well-characterized signal transduction pathways make them a favored host for gene-editing applications aimed at probing immune cell biology. By introducing ARL3 disruption into this background, researchers gain a platform to explore ciliary and potentially extra-ciliary functions of ARL3 in a lymphoid environment.
ARL3 encodes a small ADP-ribosylation factor-like GTPase that functions as a critical molecular switch in ciliary trafficking and ciliogenesis. Upon activation by the guanine nucleotide exchange factor ARL13B, GTP-bound ARL3 undergoes a conformational change that releases lipidated cargo??such as transducin and NPHP3??from the prenyl-binding proteins PDE6D and UNC119A/B, facilitating their delivery to the primary cilium. The cycle is terminated by the GTPase-activating protein RP2, which stimulates GTP hydrolysis, returning ARL3 to its inactive GDP-bound state. ARL3 also interacts with centrosomal and ciliary components including CEP164 and INPP5E, thereby coordinating intraflagellar transport, Hedgehog signaling, and the trafficking of G protein-coupled receptors to the ciliary membrane.
Although Jurkat cells lack primary cilia, the expression of ARL3 and its interacting partners in lymphoid cells raises the possibility of non-ciliary functions, such as modulation of endosomal trafficking or signal transduction. This knockout model thus enables dissection of both canonical ciliary roles??through ectopic ciliogenesis or reconstitution assays??and putative extra-ciliary activities that may impact T-cell activation, proliferation, or cytokine production. Moreover, the association of ARL3 mutations with ciliopathies like Joubert syndrome and Leber congenital amaurosis underscores the translational relevance of studying ARL3 loss-of-function in a human cell background.
Researchers can employ these ARL3 knockout cells for a wide range of downstream analyses. For example, co-immunoprecipitation and GTPase activity assays allow direct assessment of ARL3 interaction with PDE6D, UNC119, and RP2, while ciliary trafficking experiments??potentially following serum-starvation-induced ciliogenesis in Jurkat-derived ciliated models??enable quantification of cargo mislocalization. RT-qPCR and Western blotting provide routine verification of target gene disruption, and flow cytometry or immunofluorescence facilitates phenotypic characterization of immune activation markers. The polyclonal population is particularly suited for small-molecule screening campaigns aimed at identifying pharmacological modulators of ARL3 activity or ciliary transport, as well as for CRISPR-based pooled screens. For additional information about this knockout model, please contact Ascent Research.
