Microinjection of mRNA into Somatic Cells

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

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.

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.

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References

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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]