Stem cells and related proteins are the focus of many scientific studies. The ability to label specific proteins within stem cells may provide important information about protein dynamics and function. Here we use a technology to covalently tether different wavelength fluorophores to a specially designed reporter, the HaloTag® protein. The HaloTag® protein, either alone or fused to a protein of interest, was expressed in human neural stem cells (hNSC) and successfully labeled with several HaloTag® Ligands. The different colored cell-impermeant and cell-permeant fluorophores showed spatial separation and real-time translocation of membrane and internal protein pools in live hNSC. In addition, the HaloTag® protein continued to be expressed after hNSC differentiation into neurons and astrocytes. Thus, the HaloTag® technology can be used to study proteins of interest, and as such, represents a new research tool to further reveal protein biology in stem cells.
Promega Notes 95, 20–22.
Soshana Svendsen1, Erin McMillan2, Chad Zimprich1, Clive N. Svendsen2, Georgyi V. Los1