Tài liệu Báo cáo khoa học: Bioimaging of the unbalanced expression of microRNA9 and microRNA9*

Bioimaging of the unbalanced expression of microRNA9

and microRNA9* during the neuronal differentiation of

P19 cells

Mee Hyang Ko1,2,3, Soonhag Kim2,4,*, Do Won Hwang1,2,3, Hae Young Ko2,3, Young Ha Kim2,3 and

Dong Soo Lee2,4,*

1 Programs in Neuroscience, Seoul National University, Korea

2 Department of Nuclear Medicine, Seoul National University College of Medicine, Korea

3 Laboratory of Molecular Imaging and Therapy of Cancer Research Institute, Seoul National University College of Medicine, Korea

4 Medical Research Center, Seoul National University College of Medicine, Korea

MicroRNAs (miRs) are a class of small non-coding

RNA molecules, encoded as short inverted repeats in

the genomes of plants and animals. miRs are believed

to modulate the post-transcriptional regulations of

their targets in diverse biological regulatory systems

including cellular development [1,2], cell differentiation

[3], fat metabolism [4], cell proliferation and cell death

[5]. Hundreds of miRs have been isolated from mam￾malian species and a dozen of these, including

miR124a, miR9, miR128, miR131, miR178 and

Keywords

bioimaging; Luciferase; microRNA;

microRNA9 and microRNA9*; neurogenesis

Correspondence

S. Kim, Department of Nuclear Medicine,

Medical Research Center, Seoul National

University College of Medicine, 28 Yongon￾dong, Jongno-gu, Seoul 110 744, Korea

Fax: +82 2 3668 7090

Tel: +82 2 3668 7028

E-mail: [email protected]

D. S. Lee, Department of Nuclear Medicine,

Medical Research Center, Seoul National

University College of Medicine, 28 Yongon￾dong, Jongno-gu, Seoul 110 744, Korea

Fax: +82 2 3668 7090

Tel: +82 2 2072 2501

E-mail: [email protected]

*These authors contributed equally to this

work

(Received 18 January 2008, revised 10

March 2008, accepted 17 March 2008)

doi:10.1111/j.1742-4658.2008.06408.x

Generally, the 3¢-end of the duplex microRNA (miR) precursor (pre-miR)

is known to be stable in vivo and serve as a mature form of miR. However,

both the 3¢-end (miR9) and 5¢-end (miR9*) of a brain-specific miR9 have

been shown to function biologically in brain development. In this study,

real-time PCR analysis and in vitro ⁄ in vivo bioluminescent imaging demon￾strated that the upstream region of a primary miR9-1 (pri-miR9-1) can be

used to monitor the highly expressed pattern of endogenous pri-miR9-1

during neurogenesis, and that the Luciferase reporter gene can image the

unequal expression patterns of miR9 and miR9* seen during the neuronal

differentiation of P19 cells. This demonstrates that our bioimaging system

can be used to study the participation of miRs in the regulation of neur￾onal differentiation.

Abbreviations

Dicer, RNase III endonuclease; FLuc, Firefly Luciferase; GLuc, Gaussia Luciferase; miR, microRNA; miR9*, microRNA9*, 5¢-end of pre-miR9;

miR9, microRNA9, 3¢-end of miR9 precursor; piRNA, Piwi-interacting RNA; pre-miR, precursor microRNA; pre-miR9, miR9 precursor or

precursor miR9; pri-miR9-1, primary miR9-1; ROI, region of interest analysis.

FEBS Journal 275 (2008) 2605–2616 ª 2008 The Authors Journal compilation ª 2008 FEBS 2605