Plasmids encoding reprogramming factors and plasmids encoding gene-editing machinery are transfected together into fibroblast cells. Approximately three weeks later, induced pluripotent stem cell (iPSC) colonies grown from single cells are apparent. These clones can be individually picked from the dish for further isolated growth and study.
See full infographic: WEB© GEORGE RETSECK
In theory, mutating a gene of interest inside stem cells enables researchers to analyze the effects of that mutation on the development of particular cell types. In the laboratory of Jack Parent at the University of Michigan Medical School, for example, postdoctoral researcher Andrew Tidball is using such an approach to investigate how gene mutations associated with epileptic encephalopathy affect brain cell development. But while trying to introduce the specific mutations into human induced pluripotent stem cells (iPSCs), he ran into difficulties.
A major problem, Tidball says, is that after transfecting iPSCs with gene-editing plasmids, individual cells need to be isolated, but “stem cells don’t like to be [alone]. They die unless you add some components to help them along.” Even then, he adds, “a ...
