LAMTOR4 Knockout HAP1 Cell Line

The choice depends on whether you are studying LAMTOR4 (Ragulator subunit p10, C7orf59)'s role as a Ragulator complex subunit or its functions in lysosomal mTORC1 recruitment. The Knockout line is the standard tool for asking whether LAMTOR4 is required for amino acid-induced mTORC1 activation — the Ragulator complex (LAMTOR1-5, with LAMTOR1/p18 anchored at lysosomes) recruits Rag GTPases to lysosomes, which in turn recruit mTORC1 for amino acid-activated kinase activity. Overexpression is useful for studying LAMTOR4 in heterologous expression contexts. For mTOR signaling research, the EDITGENE LAMTOR4 Knockout in HAP1 enables study of Ragulator-Rag-mTORC1 signaling. Rescue with wild-type LAMTOR4 enables structure-function studies. The knockout is valuable for studying amino acid sensing pathways, mTORC1 regulation, and emerging mTORC1-targeted therapeutics — Ragulator/Rag pathway is a critical upstream regulator distinct from PI3K-Akt-TSC1/2 inputs.

Primary applications: • Amino acid-induced mTORC1 activation: phospho-S6K (T389), phospho-4E-BP1, and phospho-S6 following amino acid stimulation in LAMTOR4-null cells. • Lysosomal mTORC1 recruitment: confocal imaging of mTOR-LAMP1 colocalization following amino acid stimulation. • Ragulator complex stability: co-immunoprecipitation analysis of Ragulator (LAMTOR1-5) and Rag GTPase complex integrity. • Autophagy regulation: LC3-II accumulation and autophagic flux given mTORC1's role in autophagy inhibition. EDITGENE recommends this model for researchers investigating amino acid sensing, Ragulator-Rag-mTORC1 signaling, and lysosomal mTORC1 regulation.

Yes. LAMTOR4 rescue experiments require attention to Ragulator complex integrity: • Construct design: use a codon-modified LAMTOR4 sequence with a small C-terminal tag (FLAG, HA). LAMTOR4 is a small (~10 kDa) Ragulator complex subunit — preserve protein integrity. • Ragulator partnership: LAMTOR4 requires other Ragulator subunits (LAMTOR1, LAMTOR2, LAMTOR3, LAMTOR5) for complex formation — rescue interpretation considers Ragulator complex integrity. • Functional readout: rescue should restore amino acid-induced phospho-S6K (T389) and mTORC1 lysosomal recruitment. HAP1-specific considerations: • Diploidization: HAP1 cells gradually diploidize during extended culture — confirm ploidy by flow cytometry at the time of phenotypic assay. • Integration site sensitivity: position effects on transgene expression are more pronounced in near-haploid backgrounds; generating multiple independent rescue clones is strongly recommended. • Transduction efficiency: HAP1 transduces with lentivirus at moderate efficiency — increase MOI compared to standard immortalized lines.