TY - JOUR
T1 - Integration of CpG-free DNA induces de novo methylation of CpG islands in pluripotent stem cells
AU - Takahashi, Yuta
AU - Wu, Jun
AU - Suzuki, Keiichiro
AU - Martinez-Redondo, Paloma
AU - Li, Mo
AU - Liao, Hsin Kai
AU - Wu, Min Zu
AU - Hernández-Benítez, Reyna
AU - Hishida, Tomoaki
AU - Shokhirev, Maxim Nikolaievich
AU - Rodriguez Esteban, Concepcion
AU - Sancho-Martinez, Ignacio
AU - Izpisua Belmonte, Juan Carlos
N1 - Funding Information:
We thank M. C. Ku and T. Nguyen for next-generation sequencing; I. Dubova and K. McIntyre for teratoma analysis; D. O'Keefe for critical reading of the manuscript; C. J. Chang, A. Goebl, E. Aizawa, N. Y. Kim, Y. Hishida, R. D. Soligalla, and M. Morales Valencia for experimental help; M. Schwarz and P. Schwarz for administrative help; and A. Fukamizu for scientific discussion. Y.T. was partially supported by Astellas Foundation, Uehara Memorial Foundation, and Mochida Memorial Foundation research fellowships. K.S. and M.L. were supported by a California Institute for Regenerative Medicine (CIRM) Training Grant fellowship. P.M.-R. was partially supported by the Fundaci?n Alfonso Martin Escudero. This work was supported by the Razavi Newman Integrative Genomics and Bioinformatics Core Facility and the Next Generation Sequencing (NGS) Core Facility at the Salk Institute, with funding from NIH-National Cancer Institute (NCI) Cancer Center Support Grant (CCSG) no. P30 014195, the Chapman Foundation, and The Leona M. and Harry B. Helmsley Charitable Trust. Work in the laboratory of J.C.I.B. was supported by UCAM (iPSC work), The G. Harold and Leila Y. Mathers Charitable Foundation, The Moxie Foundation, and The Leona M. and Harry B. Helmsley Charitable Trust.
PY - 2017/5/5
Y1 - 2017/5/5
N2 - CpG islands (CGIs) are primarily promoter-associated genomic regions and are mostly unmethylated within highly methylated mammalian genomes. The mechanisms by which CGIs are protected from de novo methylation remain elusive. Here we show that insertion of CpG-free DNA into targeted CGIs induces de novo methylation of the entire CGI in human pluripotent stem cells (PSCs). The methylation status is stably maintained even after CpG-free DNA removal, extensive passaging, and differentiation. By targeting the DNA mismatch repair gene MLH1 CGI, we could generate a PSC model of a cancer-related epimutation. Furthermore, we successfully corrected aberrant imprinting in induced PSCs derived from an Angelman syndrome patient. Our results provide insights into how CpG-free DNA induces de novo CGI methylation and broaden the application of targeted epigenome editing for a better understanding of human development and disease.
AB - CpG islands (CGIs) are primarily promoter-associated genomic regions and are mostly unmethylated within highly methylated mammalian genomes. The mechanisms by which CGIs are protected from de novo methylation remain elusive. Here we show that insertion of CpG-free DNA into targeted CGIs induces de novo methylation of the entire CGI in human pluripotent stem cells (PSCs). The methylation status is stably maintained even after CpG-free DNA removal, extensive passaging, and differentiation. By targeting the DNA mismatch repair gene MLH1 CGI, we could generate a PSC model of a cancer-related epimutation. Furthermore, we successfully corrected aberrant imprinting in induced PSCs derived from an Angelman syndrome patient. Our results provide insights into how CpG-free DNA induces de novo CGI methylation and broaden the application of targeted epigenome editing for a better understanding of human development and disease.
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U2 - 10.1126/science.aag3260
DO - 10.1126/science.aag3260
M3 - Article
C2 - 28473583
AN - SCOPUS:85018771317
VL - 356
SP - 503
EP - 508
JO - Science
JF - Science
SN - 0036-8075
IS - 6337
ER -