The development of microalgal biofuels is crucial for advancing the energy transition, alleviating food pressure, preserving the natural environment, and addressing climate change. Despite significant global research and investment, the industry faces challenges due to high costs. However, advancements in new strains, technologies, and equipment have re-evaluated the feasibility of large-scale production. This review highlights the importance of selecting highly adaptable microalgae strains, particularly those capable of utilizing high concentrations of inorganic carbon sources and possessing stress resistance. Omics technologies and genetic editing have enhanced lipid accumulation, but risks associated with outdoor cultivation limit their feasibility. Photobioreactors (PBRs) offer a more controllable cultivation method, and optimizing their performance can further enhance lipid accumulation. Future research should focus on genetic engineering to reduce costs and improve economic viability, while also addressing the limitations of PBRs and exploring new cultivation systems. The integration of omics resources and genetic engineering tools will be essential for realizing economically viable microalgal-based biofuels.The development of microalgal biofuels is crucial for advancing the energy transition, alleviating food pressure, preserving the natural environment, and addressing climate change. Despite significant global research and investment, the industry faces challenges due to high costs. However, advancements in new strains, technologies, and equipment have re-evaluated the feasibility of large-scale production. This review highlights the importance of selecting highly adaptable microalgae strains, particularly those capable of utilizing high concentrations of inorganic carbon sources and possessing stress resistance. Omics technologies and genetic editing have enhanced lipid accumulation, but risks associated with outdoor cultivation limit their feasibility. Photobioreactors (PBRs) offer a more controllable cultivation method, and optimizing their performance can further enhance lipid accumulation. Future research should focus on genetic engineering to reduce costs and improve economic viability, while also addressing the limitations of PBRs and exploring new cultivation systems. The integration of omics resources and genetic engineering tools will be essential for realizing economically viable microalgal-based biofuels.