Vaccination is the first line of defense against influenza infections, yet influenza vaccine production methods are slow, antiquated, and expensive to effectively reduce the virus´ burden during epidemic or pandemic periods. There is a great need for alternative influenza vaccines and vaccination methods with a global scale impact. We demonstrate a strategy to generate influenza A virus in vivo using bacmid DNAs. Compared to the classical reverse genetics system, the "eight-in-one" bacmids (bcmd-RGFlu) showed higher efficiency of virus rescue in various cell types. Using a transfection-based inoculation (TBI) system, intranasal delivery in DBA/2J and Balb/c mice of bcmd-RGFlu plus 293T cells lead to the generation of lethal PR8 virus in vivo. A prime-boost intranasal vaccination strategy using TBI in the context of a bcmd-RGFlu encoding a temperature sensitive H1N1 virus resulted in protection of mice against lethal challenge with the PR8 strain. Taken together, these studies are proof-of-principle to highlight the potential of vaccination against influenza by in vivo reverse genetics.