Yamaguchi, Tomoyuki Hamanaka, Sanae Kamiya, Akihide Okabe, Motohito Kawarai, Mami Wakiyama, Yukiko Umino, Ayumi Hayama, Tomonari Sato, Hideyuki Lee, Youn-Su Kato-Itoh, Megumi Masaki, Hideki Kobayashi, Toshihiro Yamazaki, Satoshi Nakauchi, Hiromitsu Development of an All-in-One Inducible Lentiviral Vector for Gene Specific Analysis of Reprogramming <div><p>Fair comparison of reprogramming efficiencies and <em>in vitro</em> differentiation capabilities among induced pluripotent stem cell (iPSC) lines has been hampered by the cellular and genetic heterogeneity of de novo infected somatic cells. In order to address this problem, we constructed a single cassette all-in-one inducible lentiviral vector (Ai-LV) for the expression of three reprogramming factors (<em>Oct3/4</em>, <em>Klf4</em> and <em>Sox2</em>). To obtain multiple types of somatic cells having the same genetic background, we generated reprogrammable chimeric mice using iPSCs derived from Ai-LV infected somatic cells. Then, hepatic cells, hematopoietic cells and fibroblasts were isolated at different developmental stages from the chimeric mice, and reprogrammed again to generate 2nd iPSCs. The results revealed that somatic cells, especially fetal hepatoblasts were reprogrammed 1200 times more efficiently than adult hepatocytes with maximum reprogramming efficiency reaching 12.5%. However, we found that forced expression of <em>c-Myc</em> compensated for the reduced reprogramming efficiency in aged somatic cells without affecting cell proliferation. All these findings suggest that the Ai-LV system enables us to generate a panel of iPSC clones derived from various tissues with the same genetic background, and thus provides an invaluable tool for iPSC research.</p> </div> all-in-one;inducible;lentiviral;vector;Reprogramming 2012-07-18
    https://plos.figshare.com/articles/dataset/Development_of_an_All_in_One_Inducible_Lentiviral_Vector_for_Gene_Specific_Analysis_of_Reprogramming/122576
10.1371/journal.pone.0041007