Influenza virus causes epidemics and sporadic pandemics resulting in morbidity, mortality and economic losses. Influenza viruses require host genes to replicate. RNA interference (RNAi) screens can identify host genes coopted by influenza for replication. Targeting these pro-influenza genes can provide therapeutic strategies to reduce virus replication. Using human lung (A549) cells, 19 pro-influenza GPCR and 13 pro-influenza ion channel genes were identified using small inferring RNAs (siRNA). These pro-influenza genes were authenticated by testing A/WSN/33, A/CA/04/09, and B/Yamagata/16/1988-infected A549 cells resulting in 16 pro-influenza GPCR and 5 pro-influenza ion channel genes being validated. These findings showed that several GPCR and ion channel genes are needed for production of infectious influenza virus. These data provide potential targets for the development of host-directed therapeutic strategies to impede the influenza productive cycle to limit infection.IMPORTANCE: Influenza epidemics result in morbidity and mortality each year. Vaccines are the most effective preventive measure but require annual reformulation as mismatch of vaccine strains can result in vaccine failure. Antiviral measures are desirable particularly when vaccines fail. In this study, we used RNAi screening to identify several GPCR and ion channel genes needed for influenza virus replication. Understanding the host genes usurped by influenza during viral replication can help identify host genes that can be targeted for drug repurposing or for the development of antiviral drugs. Targeting host genes is also refractory to drug resistance generated by viral mutations, as well as provides a platform for the development of broad spectrum anti-viral drugs.