Generation and characterization of induced pluripotent stem cells from Aid-deficient mice.
It has been shown that DNA demethylation plays a pivotal role in the generation of induced pluripotent stem (iPS) cells. However, the underlying mechanism of this action is still unclear. Previous reports indicated that activation-induced cytidine deaminase (Aid, also known as Aicda) is involved in DNA demethylation in several developmental processes, as well as cell fusion-mediated reprogramming. Based on these reports, we hypothesized that Aid may be involved in the DNA demethylation that occurs during the generation of iPS cells. In this study, we examined the function of Aid in iPS cell generation using Aid knockout (Aid⁻/⁻) mice expressing a GFP reporter under the control of a pluripotent stem cell marker, Nanog. By introducing Oct3/4, Sox2, Klf4 and c-Myc, Nanog-GFP-positive iPS cells could be generated from the fibroblasts and primary B cells of Aid⁻/⁻ mice. Their induction efficiency was similar to that of wild-type (Aid⁺/⁺) iPS cells. The Aid⁻/⁻ iPS cells showed normal proliferation and gave rise to chimeras, indicating their capacity for self-renewal and pluripotency. A comprehensive DNA methylation analysis showed only a few differences between Aid⁺/⁺ and Aid⁻/⁻ iPS cells. These data suggest that Aid does not have crucial functions in DNA demethylation during iPS cell generation.
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Ren
R
Shimamoto
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Naoki
N
Amano
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Tomoko
T
Ichisaka
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Akira
A
Watanabe
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan; Gladstone Institute of Cardiovascular Disease, San Francisco, California, United States of America.
Shinya
S
Yamanaka
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Keisuke
K
Okita
D000818:Animals; D003564:Cytidine Deaminase / Q000172:deficiency* / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D000090062:Kruppel-Like Factor 4; D008810:Mice, Inbred C57BL; D000098830:AICDA (Activation-Induced Cytidine Deaminase)
C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D000098830:AICDA (Activation-Induced Cytidine Deaminase); D003564:Cytidine Deaminase