Genetically Engineered Cell Lines for TSC Research

Human embryonic kidney HEK293T cells with or without somatic genomic deletion (KO) of TSC2 using CRISPR–Cas9 genome editing were a kind gift of the TSC Alliance and Dr Nellist [26 (link)]. TRI102 cells derived from a TSC2‐null human AML and TRI103 [27 (link), 28 (link)] cells derived from TRI102 cells stably transfected with wild‐type TSC2 (pcDNA3.1 TSC2‐zeo) were obtained from the ATCC (Manassas, VI, USA). RCC cell lines from human tRCC (PRCC–TFE3; UOK124) and sporadic clear cell RCC (ccRCC; UOK111) were a kind gift from Dr W Marston Linehan [22 (link)]). All cell lines were maintained in DMEM high glucose medium (Gibco/Thermo Fisher Scientific, Waltham, MA, USA) with l‐glutamine, 10% FBS, and 1% penicillin/streptomycin stock solution at 37 °C in 5% CO₂. RCC cell lines were cultured as above in DMEM containing additional 1X essential amino acids. Rapamycin (200 nm) and Torin 1 (1 μm) (LC Laboratories, Woburn, MA, USA) treatment was performed for 72 h prior to cell lysis for immunoblotting.