An efficient method of generating transgenic mice by pronuclear microinjection.

Genetic transformation of mice using pronuclear microinjection was demonstrated by a number of groups in rapid succession in the early 1980's. Since that time, studies using transgenic animals have produced major advances in biomedical sciences and molecular genetics. More important, it is possible to study the molecular basis for tissue and stage-specific expression of genes. We recently used this method to generate transgenic mice. DNA fragment (transgene) was injected into the pronucleus of one-cell embryos. We describe this simplified protocol, which is reliable. With the use of buffered medium M2 for the whole process, it is not mandatory to have a CO2 incubator.