{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T21:33:56Z","timestamp":1791322436910,"version":"4.3.1"},"reference-count":28,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5527","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[2001,7,6]]},"abstract":"<jats:p>Cloning by nuclear transfer (NT) is an inefficient process in which most clones die before birth and survivors often display growth abnormalities. In an effort to correlate gene expression with survival and fetal overgrowth, we have examined imprinted gene expression in both mice cloned by nuclear transfer and in the embryonic stem (ES) cell donor populations from which they were derived. The epigenetic state of the ES cell genome was found to be extremely unstable. Similarly, variation in imprinted gene expression was observed in most cloned mice, even in those derived from ES cells of the same subclone. Many of the animals survived to adulthood despite widespread gene dysregulation, indicating that mammalian development may be rather tolerant to epigenetic aberrations of the genome. These data imply that even apparently normal cloned animals may have subtle abnormalities in gene expression.<\/jats:p>","DOI":"10.1126\/science.1061402","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:39:52Z","timestamp":1027762792000},"page":"95-97","source":"Crossref","is-referenced-by-count":583,"title":["Epigenetic Instability in ES Cells and Cloned Mice"],"prefix":"10.1126","volume":"293","author":[{"given":"David","family":"Humpherys","sequence":"first","affiliation":[{"name":"Whitehead Institute for Biomedical Research,"},{"name":"Department of Biology, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge MA 02142, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin","family":"Eggan","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research,"},{"name":"Department of Biology, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge MA 02142, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidenori","family":"Akutsu","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI 96822, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Konrad","family":"Hochedlinger","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research,"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"III","given":"William M.","family":"Rideout","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research,"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Detlev","family":"Biniszkiewicz","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research,"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryuzo","family":"Yanagimachi","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Reproductive Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI 96822, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rudolf","family":"Jaenisch","sequence":"additional","affiliation":[{"name":"Whitehead Institute for Biomedical Research,"},{"name":"Department of Biology, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge MA 02142, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/385810a0"},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.282.5396.2095"},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/8632"},{"key":"e_1_3_1_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.289.5482.1188"},{"key":"e_1_3_1_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/35024082"},{"key":"e_1_3_1_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/28615"},{"key":"e_1_3_1_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/9632"},{"key":"e_1_3_1_9_2","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod64.1.44"},{"key":"e_1_3_1_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0093-691X(99)00089-8"},{"key":"e_1_3_1_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.101118898"},{"key":"e_1_3_1_12_2","doi-asserted-by":"publisher","DOI":"10.1530\/ror.0.0030155"},{"key":"e_1_3_1_13_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.genet.31.1.493"},{"key":"e_1_3_1_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/35047554"},{"key":"e_1_3_1_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0093-691X(99)00263-0"},{"key":"e_1_3_1_16_2","doi-asserted-by":"publisher","DOI":"10.1071\/RD98056"},{"key":"e_1_3_1_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/72753"},{"key":"e_1_3_1_18_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.26.14984"},{"key":"e_1_3_1_19_2","unstructured":"Total RNA was isolated using TriReagent (Molecular Research Center Cincinnati OH). Five to ten micrograms were resolved in 1% agarose\u2013formaldehyde gels and transferred to Zetabind (Cuno Meridian CT) or Genescreen (NEN Boston MA) filters which were then hybridized with probe in Church buffer overnight. The probes were generated by random hexamer labeling of the following sources: H19 generated from a Bam HI\u2013Stu I 754\u2013base pair (bp) genomic fragment Peg1 \/ Mest was amplified from cDNA with primers 5\u2032-CCAAAAGCTCCTCAAAGACG-3\u2032 and 5\u2032-TCTGTTAGAAGCAGCCGATG-3\u2032 followed by nested primers 5\u2032-CTTTCCCCCATCCTCACC-3\u2032 and 5\u2032-CAAGCTCCTCCCTCACTCTG-3\u2032; Meg1 \/ Grb10 was generated from IMAGE clone #746564 and Igf2 was generated from a 1.2-kb cDNA fragment."},{"key":"e_1_3_1_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/375034a0"},{"key":"e_1_3_1_21_2","doi-asserted-by":"publisher","DOI":"10.1038\/ng0495-407"},{"key":"e_1_3_1_22_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.10.8.1008"},{"key":"e_1_3_1_23_2","unstructured":"Web figures 1 and 2 are available at Science Online at www.sciencemag.org\/cgi\/content\/full\/293\/5527\/95\/DC1."},{"key":"e_1_3_1_24_2","doi-asserted-by":"publisher","DOI":"10.1242\/dev.125.12.2273"},{"key":"e_1_3_1_25_2","unstructured":"Subclones of the V6.5 ES cell line were obtained as part of the targeting of a tet transactivator vector into the ROSA26 locus (16). Individual targeted subclones were derived by picking and expanding drug resistant colonies. Derivation culture and targeted mutagenesis of ES cells were carried out as previously described (16)."},{"key":"e_1_3_1_26_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.18.8424"},{"key":"e_1_3_1_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0925-4773(97)00655-2"},{"key":"e_1_3_1_28_2","doi-asserted-by":"publisher","DOI":"10.1242\/dev.110.3.815"},{"key":"e_1_3_1_29_2","unstructured":"We thank A. Wutz for providing subclones of the V6.5 cell line. Supported by the Victoria and Bradley Geist Foundation the Kosasa Family Foundation and the Harold Castle Foundation (R.Y.) and by NIH grants 5-R35-CA44339 and RO1-CA84198 (R.J.)."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.1061402","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,9]],"date-time":"2024-01-09T22:24:54Z","timestamp":1704839094000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.1061402"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,7,6]]},"references-count":28,"journal-issue":{"issue":"5527","published-print":{"date-parts":[[2001,7,6]]}},"alternative-id":["10.1126\/science.1061402"],"URL":"https:\/\/doi.org\/10.1126\/science.1061402","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2001,7,6]]}}}