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

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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.

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References

  1. Wong BY, Chen H, Chung SW et al (1994) High-efficiency identification of genes by functional analysis from a retroviral cDNA expression library. J Virol 68:5523–5531

    PubMed  CAS  Google Scholar 

  2. Kassner PD (2008) Discovery of novel targets with high throughput RNA interference screening. Comb Chem High Throughput Screen 11:175–188. doi:10.2174/138620708783877744

    Article  PubMed  CAS  Google Scholar 

  3. Caldwell JS (2007) Cancer cell-based genomic and small molecule screens. Adv Cancer Res 96:145–173. doi:10.1016/S0065-230X(06)96006-0

    Article  PubMed  CAS  Google Scholar 

  4. Jones AK, Buckingham SD, Sattelle DB (2005) Chemistry-to-gene screens in Caenorhabditis elegans. Nat Rev Drug Discov 4:321–330. doi:10.1038/nrd1692

    Article  PubMed  CAS  Google Scholar 

  5. Takahashi K, Tanabe K, Ohnuki M et al (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131:861–872. doi:10.1016/j.cell.2007.11.019

    Article  PubMed  CAS  Google Scholar 

  6. Nakagawa M, Koyanagi M, Tanabe K et al (2008) Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat Biotechnol 26:101–106. doi:10.1038/nbt1374

    Article  PubMed  CAS  Google Scholar 

  7. Yu J, Vodyanik MA, Smuga-Otto K et al (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science 318:1917–1920. doi:10.1126/science.1151526

    Article  PubMed  CAS  Google Scholar 

  8. Kurita R, Sasaki E, Yokoo T et al (2006) Tal1/Scl gene transduction using a lentiviral vector stimulates highly efficient hematopoietic cell differentiation from common marmoset (Callithrix jacchus) embryonic stem cells. Stem Cells 24:2014–2022. doi:10.1634/stemcells.2005-0499

    Article  PubMed  CAS  Google Scholar 

  9. Kyba M, Perlingeiro RC, Daley GQ (2002) HoxB4 confers definitive lymphoid-myeloid engraftment potential on embryonic stem cell and yolk sac hematopoietic progenitors. Cell 109:29–37. doi:10.1016/S0092-8674(02)00680-3

    Article  PubMed  CAS  Google Scholar 

  10. Wang Y, Yates F, Naveiras O et al (2005) Embryonic stem cell-derived hematopoietic stem cells. Proc Natl Acad Sci USA 102:19081–19086. doi:10.1073/pnas.0506127102

    Article  PubMed  CAS  Google Scholar 

  11. Rayner JR, Gonda TJ (1994) A simple and efficient procedure for generating stable expression libraries by cDNA cloning in a retroviral vector. Mol Cell Biol 14:880–887

    PubMed  CAS  Google Scholar 

  12. Whitehead I, Kirk H, Kay R (1995) Expression cloning of oncogenes by retroviral transfer of cDNA libraries. Mol Cell Biol 15:704–710

    PubMed  CAS  Google Scholar 

  13. Kitamura T (1998) New experimental approaches in retrovirus-mediated expression screening. Int J Hematol 67:351–359. doi:10.1016/S0925-5710(98)00025-5

    Article  PubMed  CAS  Google Scholar 

  14. Hacein-Bey-Abina S, Von Kalle C, Schmidt M et al (2003) LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 302:415–419. doi:10.1126/science.1088547

    Article  PubMed  CAS  Google Scholar 

  15. Kawano Y, Yoshida T, Hieda K et al (2004) A lentiviral cDNA library employing lambda recombination used to clone an inhibitor of human immunodeficiency virus type 1-induced cell death. J Virol 78:11352–11359. doi:10.1128/JVI.78.20.11352-11359.2004

    Article  PubMed  CAS  Google Scholar 

  16. Bai Y, Soda Y, Izawa K et al (2003) Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector. Gene Ther 10:1446–1457. doi:10.1038/sj.gt.3302026

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Division of Molecular and Clinical Genetics, Department of Molecular Genetics, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Ryo Kurita, Tatsuo Oikawa, Michiyo Okada, Tomoko Yokoo, Yuusuke Kurihara, Yuko Honda, Rui Kageyama, Youko Suehiro, Toshihiko Okazaki & Kenzaburo Tani

  2. Department of Advanced Cell and Molecular Therapy, Kyushu University Hospital, Kyushu University, Fukuoka, Japan

    Mutsunori Iga & Kenzaburo Tani

  3. Subteam for Manipulation of Cell Fate, RIKEN BioResource Center, Tsukuba, Ibaraki, 305-0074, Japan

    Hiroyuki Miyoshi

Authors
  1. Ryo Kurita
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  2. Tatsuo Oikawa
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  3. Michiyo Okada
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  4. Tomoko Yokoo
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  5. Yuusuke Kurihara
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  6. Yuko Honda
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  7. Rui Kageyama
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  8. Youko Suehiro
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  9. Toshihiko Okazaki
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  10. Mutsunori Iga
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  11. Hiroyuki Miyoshi
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  12. Kenzaburo Tani
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Corresponding author

Correspondence to Kenzaburo Tani.

Additional information

Ryo Kurita and Tatsuo Oikawa contributed equally to this work.

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Kurita, R., Oikawa, T., Okada, M. et al. Construction of a high-performance human fetal liver-derived lentiviral cDNA library. Mol Cell Biochem 319, 181–187 (2008). https://doi.org/10.1007/s11010-008-9891-5

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  • DOI: https://doi.org/10.1007/s11010-008-9891-5

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