title: Genomic instability during reprogramming by nuclear transfer is DNA replication dependent.
abstract: Somatic cells can be reprogrammed to a pluripotent state by nuclear transfer into oocytes, yet developmental arrest often occurs. While incomplete transcriptional reprogramming is known to cause developmental failure, reprogramming also involves concurrent changes in cell cycle progression and nuclear structure. Here we study cellular reprogramming events in human and mouse nuclear transfer embryos prior to embryonic genome activation. We show that genetic instability marked by frequent chromosome segregation errors and DNA damage arise prior to, and independent of, transcriptional activity. These errors occur following transition through DNA replication and are repaired by BRCA1. In the absence of mitotic nuclear remodelling, DNA replication is delayed and errors are exacerbated in subsequent mitosis. These results demonstrate that independent of gene expression, cell-type-specific features of cell cycle progression constitute a barrier sufficient to prevent the transition from one cell type to another during reprogramming.
authors.affiliation: Department of Pediatrics, Columbia University, New York 10032, USA.
authors.forename: Gloryn
authors.initials: G
authors.lastname: Chia
authors.affiliation: Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Judith
authors.initials: J
authors.lastname: Agudo
authors.affiliation: Reproductive Medicine Associates of New Jersey, New Jersey 07960, USA.
authors.forename: Nathan
authors.initials: N
authors.lastname: Treff
authors.affiliation: Center for Women's Reproductive Care, College of Physicians and Surgeons, Columbia University, New York, New York 10019, USA.|Department of Obstetrics and Gynecology, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
authors.forename: Mark V
authors.initials: MV
authors.lastname: Sauer
authors.affiliation: Institute for Cancer Genetics, College of Physicians and Surgeons, Columbia University, New York 10032, USA.
authors.forename: David
authors.initials: D
authors.lastname: Billing
authors.affiliation: Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Brown
authors.affiliation: Institute for Cancer Genetics, College of Physicians and Surgeons, Columbia University, New York 10032, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Baer
authors.affiliation: Department of Pediatrics, Columbia University, New York 10032, USA.
authors.forename: Dieter
authors.initials: D
authors.lastname: Egli
mesh_terms: D000818:Animals; D019313:BRCA1 Protein / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D065150:Cellular Reprogramming / Q000235:genetics*; D020090:Chromosome Segregation / Q000235:genetics; D002877:Chromosomes, Human / Q000378:metabolism; D004249:DNA Damage / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D004261:DNA Replication / Q000235:genetics*; D004622:Embryo, Mammalian / Q000378:metabolism; D005260:Female; D015894:Genome, Human; D042822:Genomic Instability; D006801:Humans; D007399:Interphase / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL / Q000235:genetics; D008938:Mitosis / Q000235:genetics; D000066450:Mouse Embryonic Stem Cells; D053652:Nuclear Transfer Techniques; D014158:Transcription, Genetic
chemical_list: D019313:BRCA1 Protein