Microinjection of mRNA into Somatic Cells

Maki Murata‐Hori, University of Massachusetts Medical School, Worcester, Massachusetts, USA
Yu‐Li Wang, University of Massachusetts Medical School, Worcester, Massachusetts, USA

Published online: May 2005

DOI: 10.1038/npg.els.0002696

Microinjection of messenger RNA (mRNA) represents a convenient, effective approach to probe protein functions during specific period of the cell cycle.

Keywords: protein expression; mitosis; mRNA; microinjection

Figure 1. Mitotic normal rat kidney (NRK) cells. The cells are in prometaphase (a) and metaphase (b) In both cells, microinjection should be performed around the peripheral region of the cell (marked by asterisks) to avoid damaging the mitotic spindle by microinjection. Bar, 10 m.
Figure 2. Microinjection of mRNA coding for a nondegradable mutant of cyclin B, 90, into an NRK cell expressing aurora B-GFP. (a and b) An uninjected NRK cell expressing aurora B-GFP. (c and d) An NRK cell injected with mRNA for cyclin B 90 during prometaphase and incubated until anaphase. Bar, 10 m. In both cases, aurora B is localized at centromeres during prometaphase and metaphase (not shown). In the control cells (a and b), aurora B relocates from centromeres to the central spindle upon anaphase onset. However, in the cell microinjected with mRNA for cyclin B 90 (c and d), aurora B remains associated with centromeres during anaphase.
close
 References
    Glotzer M, Murray AW and Kirshner MW (1991) Cyclin is degraded by the ubiquitin pathway. Nature 349: 132–138.
    Holloway SL, Glotzer M, King RW and Murray AW (1993) Anaphase is initiated by proteolysis rather than by the inactivation of maturation-promoting factor. Cell 73: 1393–1402.
    Murata-Hori M, Tatsuka M and Wang YL (2002) Probing the dynamics and functions of aurora B kinase in living cells during mitosis and cytokinesis. Molecular Biology of the Cell 13: 1099–1108.
    book Wang YL (1994) "Microinjection of proteins into somatic cells". In: Celis JE (ed.) Handbook of Cell Biology San Diego: Academic Press, pp. 16–21.
    Wheatley SP, Hinchcliffe EH, Glotzer M, Hyman AA, Sluder G and Wang YL (1997) CDK1 inactivation regulates anaphase spindle dynamics and cytokinesis in vivo. Journal of Cell Biology 138: 385–393.

Related Sub-Topics:

Cell Cycle | Growth and Synthesis | Techniques in Cell Biology | Translation and Protein synthesis | Single Molecule Analysis | Techniques and Tools in Molecular Biology
Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

Article Title: Microinjection of mRNA into Somatic Cells
 
How to Cite close
Murata‐Hori, Maki, and Wang, Yu‐Li(May 2005) Microinjection of mRNA into Somatic Cells. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002696]