FLT3 Knockout Cell Line (HEK 293)

The choice depends on whether you are studying FLT3 (FMS-like tyrosine kinase 3, CD135)'s role as a hematopoietic receptor tyrosine kinase or modeling FLT3-mutant acute myeloid leukemia (AML). The Knockout line is the standard tool for asking whether FLT3 is required for these processes — FLT3 is a class III receptor tyrosine kinase (with KIT, PDGFR, FMS, FLT1) activated by FLT3 ligand (FLT3LG), driving hematopoietic stem/progenitor cell proliferation; FLT3 is among the most frequently mutated genes in AML, with ITD (internal tandem duplication, ~25% AML) and TKD (tyrosine kinase domain, ~10% AML) mutations driving constitutive activation. Overexpression of mutant FLT3 (FLT3-ITD, FLT3-D835Y) is useful for studying gain-of-function AML mutations. For AML research, the EDITGENE FLT3 Knockout in HEK293 is highly informative — HEK293 supports systematic structure-function studies of FLT3. Rescue with wild-type, FLT3-ITD, FLT3-D835Y, or kinase-dead FLT3 enables comprehensive AML disease mechanism studies. The knockout is a critical specificity control for ⭐⭐ midostaurin (Rydapt, FDA-approved for FLT3-mutant AML 2017), gilteritinib (Xospata, FDA-approved 2018), quizartinib (Vanflyta, FDA-approved 2023), crenolanib, and emerging FLT3-targeted AML drug development.

Primary applications: • FLT3 inhibitor specificity: critical genetic control for midostaurin (Rydapt), gilteritinib (Xospata), quizartinib (Vanflyta), crenolanib in AML drug development. • AML mutation modeling: rescue with FLT3-ITD (most common AML mutation), FLT3-D835Y (TKD mutation), and other FLT3 activating mutations for disease genotype-function studies. • FLT3 ligand signaling: phospho-FLT3 and downstream STAT5, MAPK, PI3K pathway analysis following FLT3LG stimulation. • ITD versus TKD distinction: differential FLT3 inhibitor sensitivity studies — midostaurin covers both, quizartinib is ITD-selective, gilteritinib covers both. EDITGENE recommends this model as the critical specificity control for FLT3-targeted AML drug development and FLT3 mutation genotype-function studies.

Yes. FLT3 rescue experiments are well-established for AML drug development: • Construct design: use a codon-modified FLT3 sequence with a small intracellular C-terminal tag (FLAG, HA). FLT3 has extracellular five Ig-like domains (FLT3LG binding), transmembrane span, juxtamembrane region, and split tyrosine kinase domain (TKD1, kinase insert, TKD2) — preserve all elements. • Surface localization validation: confirm plasma membrane FLT3 expression before functional assays. • Kinase-dead rescue: K644A mutation in the ATP-binding lysine abolishes catalytic activity. • AML mutation rescue: FLT3-ITD (internal tandem duplication in juxtamembrane region) and FLT3-D835Y (TKD activation loop) mutations enable AML genotype-function studies and FLT3 inhibitor sensitivity testing. • Functional readout: rescue should restore FLT3LG-induced phospho-FLT3 and downstream STAT5/MAPK/PI3K activation. HEK293 transduces efficiently with lentivirus and supports systematic FLT3 mutation analysis for AML drug development.