Poly(2-alkyl-2-oxazoline) (PAOx) hydrogels are tailorable synthetic materials with demonstrated biomedical applications, thanks to their excellent biocompatibility and tunable properties. However, their use as injectable hydrogels is challenging as it requires invasive surgical procedures to insert the formed hydrogel into the body due to their nonsoluble 3D network structures. Herein, we introduce cyclooctyne and azide functional side chains to poly(2-oxazoline) copolymers to induce in situ gelation using strain promoted alkyne-azide cycloaddition. The gelation occurs rapidly, within 5 min, under physiological conditions when two polymer solutions are simply mixed. The influence of several parameters, such as temperature and different aqueous solutions, and stoichiometric ratios between the two polymers on the structural properties of the resultant hydrogels have been investigated. The gel formation within tissue samples was verified by subcutaneous injection of the polymer solution into an ex vivo model. The degradation study of the hydrogels in vitro showed that the degradation rate was highly dependent on the type of media, ranging from days to a month. This result opens up the potential uses of PAOx hydrogels in attempts to achieve optimal, injectable drug delivery systems and tissue engineering.