Squalene is a naturally occurring triterpenoid that serves as a key biosynthetic precursor for steroids and hopanoids in microorganisms, plants, humans, and other animals. It possesses several valuable properties, including strong antioxidant activity, excellent skin penetration, and the ability to stimulate the immune system, making it widely useful in cosmetic, food, and pharmaceutical applications. Traditionally, sharks have been the main commercial source of squalene; however, environmental and sustainability concerns have created a demand for economical and eco-friendly alternatives. As a result, researchers are increasingly using biotechnological approaches to explore microorganisms as sustainable producers of squalene. This work highlights recent advances in metabolic and bioprocess engineering strategies employed in different microorganisms for the biotechnological production of this important compound.