Molecular and Cellular Biochemistry

, Volume 319, Issue 1–2, pp 181–187 | Cite as

Construction of a high-performance human fetal liver-derived lentiviral cDNA library

  • Ryo Kurita
  • Tatsuo Oikawa
  • Michiyo Okada
  • Tomoko Yokoo
  • Yuusuke Kurihara
  • Yuko Honda
  • Rui Kageyama
  • Youko Suehiro
  • Toshihiko Okazaki
  • Mutsunori Iga
  • Hiroyuki Miyoshi
  • Kenzaburo Tani
Article
First Online:
Received:
Accepted:

Abstract

The gene transduction method is a very powerful tool, not only in basic science but also in clinical medicine. Regenerative medicine is one field that has close connection with both basic and clinical. Recently, it has been reported that induced pluripotent stem (iPS) cells can be produced from somatic cells by a three or four gene transduction. We have also recently reported that lentiviral gene transfer of the tal1/scl gene can efficiently differentiate non-human primate common marmoset ES cells into hematopoietic cells without the support of stromal cells. In this study, we constructed a high-performance human fetal liver-derived lentiviral expression library, which contains a high number of individual clones, in order to develop a very helpful tool for understanding early hematopoiesis and/or hepatocytosis for future regenerative medicine. Our lentiviral cDNA library consisted of more than 8 × 107 individual clones, and their average insert size was >2 kb. DNA sequence analysis for each individual inserted cDNAs revealed that >60% contained the full-length protein-coding regions for many genes including cytokine receptors, cytoplasmic proteins, protein inhibitors, and nuclear factors. The transduction efficiency on 293T cells was 100% and the average size of an integrated cDNA was ~1.1 kb. These results suggest that our lentiviral human fetal liver cDNA expression library could be a very helpful tool for accelerating the discovery of novel genes that are involved in early hematopoiesis and hepatopoiesis and to make the use of iPS cells more efficient in the field of regenerative medicine.

Keywords

Lentivirus Human fetal liver cDNA expression library Hematopoiesis Hepatopoiesis 
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Copyright information

© Springer Science+Business Media, LLC. 2008

Authors and Affiliations

  • Ryo Kurita
    • 1
  • Tatsuo Oikawa
    • 1
  • Michiyo Okada
    • 1
  • Tomoko Yokoo
    • 1
  • Yuusuke Kurihara
    • 1
  • Yuko Honda
    • 1
  • Rui Kageyama
    • 1
  • Youko Suehiro
    • 1
  • Toshihiko Okazaki
    • 1
  • Mutsunori Iga
    • 2
  • Hiroyuki Miyoshi
    • 3
  • Kenzaburo Tani
    • 1
    • 2
  1. 1.Division of Molecular and Clinical Genetics, Department of Molecular GeneticsMedical Institute of Bioregulation, Kyushu UniversityFukuokaJapan
  2. 2.Department of Advanced Cell and Molecular TherapyKyushu University Hospital, Kyushu UniversityFukuokaJapan
  3. 3.Subteam for Manipulation of Cell FateRIKEN BioResource CenterTsukubaJapan

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