Chul-Yong Parka,b,1, Jungeun Kimc,d,1, Jiyeon Kweonc,d,1, Jeong Sang Sone, Jae Souk Leea,f, Jeong-Eun Yooa,f, Sung-Rae Chog, Jong-Hoon Kime, Jin-Soo Kimc,d,2, and Dong-Wook Kima,b,f,2
aDepartment of Physiology,
bSeverance Biomedical Research Institute,
fBrain Korea 21 Plus Project for Medical Science, and
gDepartment and Research Institute of Rehabilitation Medicine, Yonsei University College of Medicine, Seoul 120-752, South Korea;
cCenter for Genome Engineering, Institute for Basic Science, Seoul 151-742, South Korea;
dDepartment of Chemistry, Seoul National University, Seoul 151-742, South Korea; and
eLaboratory of Stem Cell Biology, Division of Biotechnology, College of Life Science and Biotechnology, Korea University, Seoul 136-713, South Korea
Hemophilia A, one of the most common genetic bleeding disorders, is caused by various mutations in the blood coagulation factor VIII (F8) gene. Among the genotypes that result in hemophilia A, two different types of chromosomal inversions that involve a portion of the F8 gene are most frequent, accounting for almost half of all severe hemophilia A cases. In this study, we used a transcription activator-like effector nuclease (TALEN) pair to invert a 140-kbp chromosomal segment that spans the portion of the F8 gene in human induced pluripotent stem cells (iPSCs) to create a hemophilia A model cell line. In addition, we reverted the inverted segment back to its normal orientation in the hemophilia model iPSCs using the same TALEN pair. Importantly, we detected the F8 mRNA in cells derived from the reverted iPSCs lines, but not in those derived from the clones with the inverted segment. Thus, we showed that TALENs can be used both for creating disease models associated with chromosomal rearrangements in iPSCs and for correcting genetic defects caused by chromosomal inversions. This strategy provides an iPSC-based novel therapeutic option for the treatment of hemophilia A and other genetic diseases caused by chromosomal inversions.
genome editing, CRISPR, Cas9, ZFN
1C.-Y.P., J. Kim, and J. Kweon contributed equally to this work.
2To whom correspondence may be addressed.
Author contributions: J.-S.K. and D.-W.K. designed research; C.-Y.P., J. Kim, J. Kweon, J.S.S., J.S.L., J.-E.Y., S.-R.C., and J.-H.K. performed research; and C.-Y.P., J.-S.K., and D.-W.K. wrote the paper.