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J Nucl Med. 2008; 49 (Supplement 1):67P
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Novel Approaches to Molecular Imaging

Optical and Bioluminescence

Real time monitoring biogenesis and functional targeting of miR124a during neurogenesis

Hae young Ko1, Soonhag Kim1, Do Won Hwang1, Young Ha Kim1 and Dong Soo Lee1

1 Nuclear Medicine, Seoul National University, Seoul, South Korea

268

Objectives: The function of microRNAs (miRNAs) is translational repression or mRNA cleavage of target genes by binding to 3’-untranslated regions (3’UTR) of target mRNA. Recent studies have demonstrated that some miRNA are involved in neurogenesis. In this study, we monitored the expression of miR124a, brain specific-miRNA, and investigated the molecular functions of miR124a during neurogenesis.

Methods: We constructed miR124aP379/Gluc plasmid carrying Gaussia luciferase (Gluc) under control of a miR124a promoter to monitor the primary transcript of miR124a and CMV/Gluc/3xPT_miR124a containing three copies of the perfect target sequences of mature miR124a in Gluc vector to monitor the mature miR124a. These plasmids were transfected into P19 cells (embryonal carcinoma cell) and then in vitro and in vivo visualized by Gaussia luciferase system. Neuronal differentiation of P19 cells was induced by retinoic acid treatment. We searched the predicted targets of miR124a using the bioinformatics database. Among them, the 3’UTR of chromsome14 open reading frame 24 (C14orf24) gene was confirmed to combine with miR124a by Gluc imaging.

Results: The endogenous levels of the primary transcripts and mature forms of miR124a were increased during neuronal differentiation of P19 cells. Overexpression of only miR124a into P19 cells resulted in a preneuronal gene expression pattern. Among the genes selected from the comparison of both bioinformatically predicted targets and miR124a-downregulated genes in HeLa cells, C14orf24 gene was regulated by miR124a.

Conclusions: Our noninvasive luciferease system, which was used for successful monitoring of both miRNA biogenesis and its targets, provided the imaging information on miRNA-related neurogenesis. We could expect that our system is useful for neuronal differentiation imaging derived from stem cells.





This Article
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Right arrow Email this article to a friend
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Right arrow Alert me to new issues of the journal
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Right arrow Articles by Ko, H. y.
Right arrow Articles by Lee, D. S.
PubMed
Right arrow Articles by Ko, H. y.
Right arrow Articles by Lee, D. S.