Biotechnology Letters

, Volume 37, Issue 11, pp 2333–2340 | Cite as

Bioimaging of transcriptional activity of microRNA124a during neurogenesis

  • Hae Young Ko
  • Jonghwan Lee
  • Yong Seung Lee
  • Youngsok Choi
  • Bahy A. Ali
  • Abdulaziz A. Al-Khedhairy
  • Soonhag Kim
Original Research Paper
First Online:
Received:
Accepted:

Abstract

Objectives

A special vector system was developed to monitor the in vitro and in vivo endogenous level of a primary transcript of miR124a during neuronal differentiation

Results

The upstream regions of miR124a were fused with luciferase (Gluc) and their activity was measured. During neurogenesis of P19 cells, the primary transcript level of miR124a was increased 1.5-times compared to the undifferentiated P19 cells. P19 cells grafted to nude mice exhibited the same pattern of luciferase activity in vivo as they did in vitro.

Conclusion

The expression of primary miR124a during neurogenesis was successfully imaged by in vitro and in vivo luciferase reporter gene-based method.

Keywords

Bioluminescence imaging In vivo imaging MicroRNA124a Neurogenesis Promoter 

Notes

Acknowledgments

This work was supported by the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Korean government (MEST) (No. 2011-0019270 and No. 2013R1A2A2A01068140), the Next-Generation BioGreen 21 program (#PJ010002), Rural Development Administration and a grant of the Korean Health Technology R&D Project, Ministry of Health & Welfare (HI14C3297), and the Visiting Professor Program (VPP) at King Saud University, Kingdom of Saudi Arabia.

Supporting information

Supplementary Table 1—A list of primer sequences.

Supplementary Fig. 1—In vitro expression of pri-miR124a2 during neuronal differentiation.

Supplementary Fig. 2—Analysis of miR124a promoter.

Supplementary material

10529_2015_1912_MOESM1_ESM.docx (217 kb)
Supplementary Fig. 1. In vitro expression of pri-miR124a2 during neuronal differentiation. a Schematic diagram of promoter-fused reporter gene constructs. Five different fragments of the miR124a2 upstream region were inserted into a promoterless vector, pGLuc-basic. The promoter size of each construct is indicated by the number in the name of each construct. b Promoter analysis of pri-miR124a2. Supplementary material 1 (DOCX 217 kb)
10529_2015_1912_MOESM2_ESM.docx (24 kb)
Supplementary Fig. 2. Analysis of miR124a promoter. The miR124a1 promoter (~1.1 kb of upstream) was analyzed using the JASPAR program. Taxonomy was based on vertebrate and the threshold was 85.0 points. Relative position is shown in the number. Putative Prx and USF binding sites are highlighted by green boxes. Supplementary material 2 (DOCX 23 kb)
10529_2015_1912_MOESM3_ESM.docx (21 kb)
Supplementary Table 1. A list of primer sequences. Supplementary material 3 (DOCX 20 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Hae Young Ko
    • 1
    • 2
  • Jonghwan Lee
    • 1
    • 2
  • Yong Seung Lee
    • 1
    • 2
  • Youngsok Choi
    • 3
  • Bahy A. Ali
    • 4
    • 5
  • Abdulaziz A. Al-Khedhairy
    • 6
  • Soonhag Kim
    • 1
    • 2
  1. 1.Institute for Bio-Medical Convergence, College of MedicineCatholic Kwandong UniversityGangneung-siRepublic of Korea
  2. 2.Catholic Kwandong University International St. Mary’s HospitalIncheon Metropolitan CityRepublic of Korea
  3. 3.Department of Biomedical ScienceCHA UnversitySeongnam-siRepublic of Korea
  4. 4.Al-Jeraisy DNA Research Chair, Department of Zoology, College of ScienceKing Saud UniversityRiyadhSaudi Arabia
  5. 5.Department of Nucleic Acids Research, Genetic Engineering and Biotechnology Research InstituteCity for Scientific Research and Technological ApplicationsAlexandriaEgypt
  6. 6.Department of Zoology, College of ScienceKing Saud UniversityRiyadhSaudi Arabia

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