This review highlights the transformative applications of ultrafast technologies in biotechnology, focusing on their ability to provide real-time visualization and precise manipulation of biomolecular processes. Femtosecond lasers have enhanced precision in gene editing, minimizing off-target effects, while ultrafast spectroscopy has advanced understanding of protein folding pathways, enzymatic activity, and energy transfer mechanisms. Notable findings include the identification of protein folding intermediates linked to misfolding diseases, improved insights into enzymatic catalysis through hydration studies, and the development of real-time monitoring systems for CRISPR gene editing. Imaging innovations such as pump-probe microscopy and Coherent Anti-Stokes Raman Scattering (CARS) enable high-resolution observation of cellular dynamics, intracellular signaling, and neural activity. Furthermore, attosecond spectroscopy has provided unprecedented insights into ultrafast electron dynamics and charge migration. Integrating ultrafast technologies with AI and nanotechnology has accelerated advances in diagnostics, personalized medicine, and synthetic biology, driving breakthroughs in drug discovery, targeted therapeutics, and regenerative medicine. Despite challenges such as photodamage, integration with complex biological systems, and ethical considerations, ongoing advancements in ultrafast technologies are set to revolutionize biotechnology. These innovations hold immense potential for addressing critical challenges in healthcare and life sciences, enabling transformative progress in understanding and treating complex diseases.