Abstract
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
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
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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 μm. |
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Figure 2.
Microinjection of mRNA coding for a nondegradable mutant of cyclin B, Δ90, 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 Δ90 during prometaphase and incubated until anaphase. Bar, 10 μm. 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 Δ90 (c and d), aurora B remains associated with centromeres during anaphase. |
References
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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.
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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.
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How to Cite
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]
