Linkers are short or long stretch of amino acid sequences used to create chimeric proteins. Linkers can adopt various structures in a biological environment which is a deciding factor for the functionality of a chimeric protein. It is essential to generate fully separated proteins in the chimeric protein and an effectual linker design increases the stability and activity of the protein. In this review, we summarize the types of linkers and strategies for designing linkers in chimeric protein generation. Appropriate linker design results in the creation of recombinant fusion protein which can be employed in biochemical analysis, drug targeting, pharmaceuticals, diagnostic, and industrial and biotechnological applications.