The use of genetically engineered mice has become a standard approach in order to study the physiological contribution of genes in a variety of life-science disciplines. Classical and conditional gene-targeting methods aimed at generating knock-out mice that lack gene products have been useful, but may be limited in their scope. If the gene of interest is essential for cell viability, little insight can be gained into the in vivo function of these genes. A hypomorphic approach, utilizing many of the same methods employed for traditional gene targeting, allows one to disrupt the function of genes to a lesser degree and bypass the lethality caused by many gene mutations. The purpose of this chapter is to introduce the concepts behind how hypomorphic alleles impede normal genetic function and provide information necessary to construct a targeting vector successfully for use in ES cells to generate ultimately mice with lower than normal amounts of an endogenous protein of interest.