TY - JOUR
T1 - TALEN-mediated shift of mitochondrial DNA heteroplasmy in MELAS-iPSCs with m.13513G>A mutation
AU - Yahata, Naoki
AU - Matsumoto, Yuji
AU - Omi, Minoru
AU - Yamamoto, Naoki
AU - Hata, Ryuji
N1 - Funding Information:
We would like to express our sincere gratitude to the patient and his family for their participation in our research. We are also grateful to all our coworkers, and to Dr. Tomihiko Ide (Fujita Health University) for assistance at the Institute of Joint Research. The authors also wish to express their thanks to Dr. Shinya Yamanaka for supplying reprogramming plasmids (Addgene #27077, #27078, and #27080), and to Dr. Takashi Yamamoto for supplying the Platinum Gate TALEN kit (Addgene kit #1000000043) and the Yamamoto Lab TALEN Accessory Pack including the pGL4-SSA plasmid (Addgene #1000000030). This study was supported in part by grants from the Hori Sciences and Arts Foundation (N. Yahata), grants from the Fujita Health University Research Fund to N. Yahata and R.H., and JPSP KAKENHI Grant Numbers JP16K19675 (N. Yahata) and JP26860831 (Y.M.).
Publisher Copyright:
© 2017 The Author(s).
PY - 2017/12/1
Y1 - 2017/12/1
N2 - Induced pluripotent stem cells (iPSCs) are suitable for studying mitochondrial diseases caused by mitochondrial DNA (mtDNA) mutations. Here, we generated iPSCs from a patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) with the m.13513G>A mutation. The patient's dermal fibroblasts were reprogrammed, and we established two iPSC clones with and without mutant mtDNA. Furthermore, we tried to decrease mutant mtDNA level in iPSCs using transcription activator-like effector nucleases (TALENs). We originally engineered platinum TALENs, which were transported into mitochondria, recognized the mtDNA sequence including the m.13513 position, and preferentially cleaved G13513A mutant mtDNA (G13513A-mpTALEN). The m.13513G>A heteroplasmy level in MELAS-iPSCs was decreased in the short term by transduction of G13513A-mpTALEN. Our data demonstrate that this mtDNA-targeted nuclease would be a powerful tool for changing the heteroplasmy level in heteroplasmic iPSCs, which could contribute to elucidation of the pathological mechanisms of mitochondrial diseases caused by mtDNA mutations.
AB - Induced pluripotent stem cells (iPSCs) are suitable for studying mitochondrial diseases caused by mitochondrial DNA (mtDNA) mutations. Here, we generated iPSCs from a patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) with the m.13513G>A mutation. The patient's dermal fibroblasts were reprogrammed, and we established two iPSC clones with and without mutant mtDNA. Furthermore, we tried to decrease mutant mtDNA level in iPSCs using transcription activator-like effector nucleases (TALENs). We originally engineered platinum TALENs, which were transported into mitochondria, recognized the mtDNA sequence including the m.13513 position, and preferentially cleaved G13513A mutant mtDNA (G13513A-mpTALEN). The m.13513G>A heteroplasmy level in MELAS-iPSCs was decreased in the short term by transduction of G13513A-mpTALEN. Our data demonstrate that this mtDNA-targeted nuclease would be a powerful tool for changing the heteroplasmy level in heteroplasmic iPSCs, which could contribute to elucidation of the pathological mechanisms of mitochondrial diseases caused by mtDNA mutations.
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U2 - 10.1038/s41598-017-15871-y
DO - 10.1038/s41598-017-15871-y
M3 - Article
C2 - 29138463
AN - SCOPUS:85034054837
VL - 7
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 15557
ER -