The DMXL1 Knockout Jurkat Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout population of Jurkat T lymphocytes with targeted disruption of the DMXL1 gene. This polyclonal pool provides a mixed genetic background that enables the study of gene function without clonal selection biases, offering a robust loss-of-function model for investigating the roles of DMXL1 in endolysosomal biology and signaling.
The host Jurkat cell line is an immortalized human T lymphocyte line originally derived from an acute T-cell leukemia patient. Jurkat cells are a widely used model system for studying T-cell receptor signaling, activation, apoptosis, and leukemia biology. Their well-characterized signaling networks and rapid growth make them particularly suitable for genetic perturbation studies aimed at understanding the molecular underpinnings of T-cell malignancies and immune cell function.
DMXL1 encodes a WD40 repeat-containing scaffold protein that is essential for the assembly and activity of the vacuolar H+-ATPase (V-ATPase), a multi-subunit proton pump responsible for endolysosomal acidification. DMXL1 functions downstream of nutrient and amino acid signaling inputs that converge on mTORC1, interacting with V-ATPase subunits such as ATP6V0A1 and the LAMTOR/Ragulator complex to promote lysosomal acidification. This activity facilitates Rab7-mediated autophagosome-lysosome fusion and autophagic flux, as well as mTORC1 reactivation on lysosomal surfaces. Key pathway components include DMXL1, V-ATPase, mTORC1, Rab7, LC3, and LAMP1. Disruption of DMXL1 impairs endolysosomal acidification, blocking autophagic degradation and altering mTORC1 signaling dynamics.
In Jurkat T lymphocytes, DMXL1 knockout is expected to disrupt endolysosomal acidification and autophagic flux, processes that are particularly important for T-cell receptor signaling, metabolic reprogramming, and survival. Given the reliance of leukemia cells on autophagy for stress adaptation and nutrient recycling, loss of DMXL1 may sensitize Jurkat cells to apoptotic stimuli or impair their proliferative capacity, making this knockout model a valuable tool for studying lysosomal dysfunction in T-cell leukemia and for probing mTORC1-dependent growth control.
This DMXL1 knockout polyclonal Jurkat cell population is suited for a range of research applications, including the investigation of lysosomal dysfunction in T-cell leukemia, dissection of mTORC1 regulatory mechanisms, and evaluation of autophagy defects in cancer. Researchers can monitor lysosomal acidification using LysoTracker staining, assess autophagic flux via LC3 turnover by western blot, and measure mTORC1 activity through phospho-S6K1 analysis. Additional experiments such as immunofluorescence for LAMP1 and LC3 colocalization, cell viability assays (MTS), apoptosis detection by flow cytometry, and RT-qPCR profiling of autophagy-related genes provide complementary readouts. These cells are also amenable to screening small-molecule modulators of endolysosomal trafficking or autophagy. For further technical details or custom pooled knockout formats, please contact Ascent Research.
