AKT1S1 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Jurkat T-lymphocyte line, featuring targeted disruption of the AKT1S1 gene. This loss-of-function model provides a powerful tool for dissecting the regulatory roles of AKT1S1 (PRAS40) in mTORC1 signaling and nutrient sensing. The polyclonal format captures a heterogeneous spectrum of editing events, enabling population-level analyses that reflect biological variability.
The Jurkat host cell line is an immortalized human T-cell leukemia model widely used to study T-cell receptor signaling, activation, and oncogenic transformation. Originating from acute lymphoblastic leukemia, Jurkat cells exhibit robust growth and are genetically tractable, making them well-suited for CRISPR/Cas9-mediated gene disruption. Their inherent signaling machinery, including the PI3K/AKT/mTOR axis, provides a relevant context for investigating growth control pathways.
AKT1S1 encodes a proline-rich AKT substrate that acts as a negative regulator of mTORC1. Under nutrient-depleted conditions, AKT1S1 interacts with RPTOR and mLST8 to suppress mTORC1 kinase activity. Upon growth factor stimulation, AKT phosphorylates AKT1S1 at Thr246, triggering its dissociation from RPTOR and relieving inhibition. Activated mTORC1 then phosphorylates downstream targets such as S6K1 and 4EBP1, driving protein synthesis and cell growth. AKT1S1 also modulates autophagy through mTORC1-dependent and independent regulation of ULK1.
In the Jurkat T-cell background, disruption of AKT1S1 is expected to enhance mTORC1 signaling, influencing proliferation, survival, and metabolic reprogramming. This model is particularly valuable for exploring how constitutive mTORC1 activation contributes to leukemogenesis, drug sensitivity, and autophagy dysregulation. The polyclonal nature allows observation of diverse functional outcomes, mirroring heterogeneity observed in clinical samples.
Researchers can apply these cells in diverse assays: western blotting for phospho-AKT1S1 (Thr246) or phospho-S6K1, co-immunoprecipitation of mTORC1 components, and mTOR kinase activity measurements. Autophagy flux can be quantified by monitoring LC3 turnover with lysosomal inhibitors. Cell viability and proliferation assays in the presence of mTOR inhibitors or chemotherapeutics enable drug sensitivity profiling. These applications support studies in cancer biology, insulin resistance, and autophagy. For further technical information or to discuss your experimental needs, please contact Ascent Research.
