AKT2 Knockout Cell Line (A549)

AKT (Protein Kinase B) is a family of three paralogous serine/threonine kinases (AKT1/PKBα, AKT2/PKBβ, AKT3/PKBγ) that are activated downstream of PI3K signaling — PI3K generates PIP3, which recruits AKT to the plasma membrane via its PH domain; AKT is then phosphorylated at T308 (activation loop, by PDK1) and S473 (hydrophobic motif, by mTORC2) for full activation. AKT family members share substantial substrate scope but have distinct tissue expression and functions: AKT1 is broadly expressed and important in cell survival/growth; AKT2 is enriched in insulin-sensitive tissues (liver, muscle, adipose) and central to insulin signaling; AKT3 is enriched in brain and testis with roles in neural development. Substantial paralog redundancy means that single-isoform knockouts often show modest phenotypes — paired and combination knockouts are essential for systematic functional dissection. This AKT2 Knockout in A-549 enables study of AKT2-specific functions — AKT2 is enriched in insulin-sensitive tissues (liver, muscle, adipose) and is central to insulin-mediated glucose uptake (GLUT4 translocation); AKT2 loss-of-function causes severe insulin resistance and diabetes mellitus in mice and rare human cases. Overexpression is useful for studying AKT2 gain-of-function effects. Important consideration: AKT1/AKT3 paralog expression analysis aids interpretation given substantial redundancy. This product is part of EDITGENE's complete AKT paralog dissection toolkit in A-549 (single AKT1, single AKT2, double AKT1&AKT2, double AKT1&AKT3, double AKT2&AKT3). Rescue with wild-type, kinase-dead (K181M), or constitutively active (myristoylated) AKT2 enables structure-function studies. The knockout is valuable for studying AKT2-specific insulin signaling and AKT inhibitor isoform selectivity.

Primary applications: • AKT2-specific insulin signaling: GLUT4 translocation and insulin-induced glucose uptake analysis in heterologous insulin-relevant contexts. • AKT family paralog dissection: parallel analysis with AKT1 KO and the three double KOs in A-549 (all available) for systematic paralog studies. • Cancer biology: A-549 NSCLC proliferation and survival assays. • AKT2-selective inhibitor specificity: emerging AKT2-selective inhibitor specificity testing. EDITGENE recommends this AKT2 single KO as a key component of the systematic AKT paralog dissection toolkit.

Yes. AKT2 rescue experiments are well-established for insulin signaling research: • Construct design: use a codon-modified AKT2 sequence with a small C-terminal tag (FLAG, HA). Preserve PH domain (PIP3 binding), kinase domain (T309 activation site), and C-terminal hydrophobic motif (S474). • Kinase-dead rescue: K181M ATP-binding lysine mutation abolishes catalytic activity. • Constitutively active rescue: myr-AKT2 constitutive plasma membrane localization. • Patient mutation rescue: rare AKT2 loss-of-function mutations in human insulin resistance can be modeled. • Functional readout: rescue should restore AKT2-specific insulin-induced GLUT4 translocation. A-549-specific considerations: • A-549 is a human non-small cell lung carcinoma (NSCLC) cell line widely used for lung cancer drug development. • Lentiviral transduction is supported with moderate efficiency. • A-549's PI3K-AKT pathway activity makes it a relevant context for systematic AKT paralog research.