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 Murahashi, Hiroyuki Miyoshi, Kenzaburo Tani

Research output: Contribution to journalArticle

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)181-187
Number of pages7
JournalMolecular and cellular biochemistry
Volume319
Issue number1-2
DOIs
Publication statusPublished - Dec 1 2008

Fingerprint

Gene Library
Liver
Genes
Regenerative Medicine
Induced Pluripotent Stem Cells
Complementary DNA
Hematopoiesis
Stem cells
Clone Cells
Gene transfer
Callithrix
Proteins
Cytokine Receptors
HEK293 Cells
Clinical Medicine
DNA sequences
Genetic Association Studies
Stromal Cells
Nuclear Proteins
DNA Sequence Analysis

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Kurita, R., Oikawa, T., Okada, M., Yokoo, T., Kurihara, Y., Honda, Y., ... Tani, K. (2008). Construction of a high-performance human fetal liver-derived lentiviral cDNA library. Molecular and cellular biochemistry, 319(1-2), 181-187. https://doi.org/10.1007/s11010-008-9891-5

Construction of a high-performance human fetal liver-derived lentiviral cDNA library. / Kurita, Ryo; Oikawa, Tatsuo; Okada, Michiyo; Yokoo, Tomoko; Kurihara, Yuusuke; Honda, Yuko; Kageyama, Rui; Suehiro, Youko; Okazaki, Toshihiko; Murahashi, Mutsunori; Miyoshi, Hiroyuki; Tani, Kenzaburo.

In: Molecular and cellular biochemistry, Vol. 319, No. 1-2, 01.12.2008, p. 181-187.

Research output: Contribution to journalArticle

Kurita, R, Oikawa, T, Okada, M, Yokoo, T, Kurihara, Y, Honda, Y, Kageyama, R, Suehiro, Y, Okazaki, T, Murahashi, M, Miyoshi, H & Tani, K 2008, 'Construction of a high-performance human fetal liver-derived lentiviral cDNA library', Molecular and cellular biochemistry, vol. 319, no. 1-2, pp. 181-187. https://doi.org/10.1007/s11010-008-9891-5
Kurita, Ryo ; Oikawa, Tatsuo ; Okada, Michiyo ; Yokoo, Tomoko ; Kurihara, Yuusuke ; Honda, Yuko ; Kageyama, Rui ; Suehiro, Youko ; Okazaki, Toshihiko ; Murahashi, Mutsunori ; Miyoshi, Hiroyuki ; Tani, Kenzaburo. / Construction of a high-performance human fetal liver-derived lentiviral cDNA library. In: Molecular and cellular biochemistry. 2008 ; Vol. 319, No. 1-2. pp. 181-187.
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