Description

The LYST Knockout HEK293T Cell Line is a genomically engineered cell line featuring CRISPR/Cas9-mediated disruption of the LYST gene, which encodes the lysosomal trafficking regulator protein. This stable knockout model is derived from the HEK293T background and provides a constitutive loss-of-function system for probing lysosome-related processes. Researchers can employ this cell line to study lysosomal size regulation, intracellular trafficking, and secretory functions in a highly transfectable and experimentally tractable host.

HEK293T cells are human embryonic kidney epithelial cells transformed with SV40 large T antigen, offering adherent growth, exceptional transfection efficiency, and robust recombinant protein expression. These features render the host line ideal for assays requiring transient overexpression, viral packaging, or high-throughput imaging. The genetic stability and ease of culture further support reproducible generation of knockout derivatives for long-term studies.

The LYST protein functions as a central scaffold in lysosomal biogenesis and trafficking, operating downstream of the transcription factor TFEB and the mTORC1 nutrient-sensing pathway. LYST interacts with late endosomal/lysosomal proteins LAMP1 and LAMP2, small GTPases Rab7 and Rab27A, and the SNARE VAMP7 to regulate lysosome fission, intracellular distribution, and fusion with effector organelles. Its disruption in HEK293T cells abrogates these interactions, yielding pathologically enlarged lysosomes, reduced autophagic flux, and impaired secretion of lysosomal enzymes.

The LYST knockout HEK293T model recapitulates giant lysosomes and secretory defects characteristic of Chediak-Higashi syndrome, providing a simplified system to study lysosome-related organelle pathology. It enables dissection of the interplay between lysosomal positioning, endosomal maturation, and cytotoxic granule release, and is amenable to genetic and pharmacologic rescue experiments. This cell line thus serves as a platform for investigating disease mechanisms and testing interventions targeting lysosomal dysfunction.

This knockout cell line supports a variety of experimental approaches, including high-content drug screening for lysosomal storage correctors, live-cell imaging with LysoTracker or LAMP1 reporters, ??-hexosaminidase secretion assays, and co-immunoprecipitation of LYST-associated complexes. It is also well-suited for functional genomics studies and evaluation of small molecules that modulate the TFEB?CmTORC1 axis. For additional details, please reach out to Ascent Research.