AKT1 and AKT3 Knockout A-549 Cell Line

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 AKT1 & AKT3 Double Knockout in A-549 is uniquely valuable for asking which functions are AKT1/AKT3-dependent versus AKT2-dependent — combined loss of AKT1 and AKT3 leaves only AKT2 as the functional AKT isoform, which is particularly useful for isolating AKT2-mediated insulin signaling. Single-isoform rescue (AKT1 alone or AKT3 alone) in the double knockout enables paralog-specific functional dissection. For systematic AKT family research, this product is part of EDITGENE's complete AKT paralog dissection toolkit. The double KO enables clean AKT2-only signaling analysis. The knockout is a critical specificity tool for capivasertib (Truqap), ipatasertib, MK-2206, ARQ-092, and isoform-selective emerging AKT inhibitors.

Primary applications: • AKT2-only signaling: pure AKT2-mediated insulin signaling analysis given AKT1/3 loss — ideal for studying insulin/metabolic AKT functions. • Pan-AKT inhibitor specificity: critical genetic control for capivasertib, ipatasertib in the AKT2-only background. • Single-isoform rescue: AKT1 alone or AKT3 alone re-introduction enables paralog-specific dissection. • AKT paralog matrix dissection: complete AKT paralog dissection toolkit analysis. EDITGENE recommends this double KO for systematic AKT2-only signaling research.

Yes, and rescue experiments are uniquely powerful in this AKT1&3 double knockout: • Single-isoform rescue: re-introduction of AKT1 alone or AKT3 alone enables paralog-specific dissection. • Construct design: same considerations as AKT family rescues. • AKT1 E17K rescue: cancer-associated AKT1 E17K activating mutation enables HR+ breast cancer disease modeling. • Functional readout: rescue should restore PI3K-AKT signaling. 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.