ArticleEpigenetics2011
Limiting dilution bisulfite (pyro)sequencing reveals parent-specific methylation patterns in single early mouse embryos and bovine oocytes.
Article in Epigenetics, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 76 citations in OpenAlex.
- Genome-wide assessment of DNA methylation alterations induced by superovulation, sexual immaturity and in vitro follicle growth in mouse blastocysts.Clinical epigenetics · 2023Article
- Targeted Methylation Profiling of Single Laser-Capture Microdissected Post-Mortem Brain Cells by Adapted Limiting Dilution Bisulfite Pyrosequencing (LDBSP).International journal of molecular sciences · 2022Article
- Along the Bos taurus genome, uncover candidate imprinting control regions.BMC genomics · 2022Article
- scTEM-seq: Single-cell analysis of transposable element methylation to link global epigenetic heterogeneity with transcriptional programs.Scientific reports · 2022Article
- Loss of methylation of H19-imprinted gene derived from assisted reproductive technologies can be mitigated by cleavage-stage embryo transfer in mice.Journal of assisted reproduction and genetics · 2019Article
- DNA methylation analysis and editing in single mammalian oocytes.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- Maintenance ofEnvironmental epigenetics · 2017Review
- High Frequency of Imprinted Methylation Errors in Human Preimplantation Embryos.Scientific reports · 2015Article
- Single-cell, locus-specific bisulfite sequencing (SLBS) for direct detection of epimutations in DNA methylation patterns.Nucleic acids research · 2015Article
- Multiplexed locus-specific analysis of DNA methylation in single cells.Nature protocols · 2015Article
- Genes and Conditions Controlling Mammalian Pre- and Post-implantation Embryo Development.Current genomics · 2015Article
- Bisphenol A Effects on Mammalian Oogenesis and Epigenetic Integrity of Oocytes: A Case Study Exploring Risks of Endocrine Disrupting Chemicals.BioMed research international · 2015Review
- Epigenetic heterogeneity of developmentally important genes in human sperm: implications for assisted reproduction outcome.Epigenetics · 2014Article
- Advances in the profiling of DNA modifications: cytosine methylation and beyond.Nature reviews. Genetics · 2014Review
- The specification of imprints in mammals.Heredity · 2014Review
- Epigenetic disorders and altered gene expression after use of Assisted Reproductive Technologies in domestic cattle.Epigenetics · 2014Review
- Single cell genomics: advances and future perspectives.PLoS genetics · 2014Review
- Metabolic programming of MEST DNA methylation by intrauterine exposure to gestational diabetes mellitus.Diabetes · 2013Article
- Constitutive promoter methylation of BRCA1 and RAD51C in patients with familial ovarian cancer and early-onset sporadic breast cancer.Human molecular genetics · 2012Article
- Single oocyte bisulfite mutagenesis.Journal of visualized experiments : JoVE · 2012Article
Corrections and comments
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Authors and funding
13 authors at 5 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
To detect rare epigenetic effects associated with assisted reproduction, it is necessary to monitor methylation patterns of developmentally important genes in a few germ cells and individual embryos. Bisulfite treatment degrades DNA and reduces its complexity, rendering methylation analysis from small amounts of DNA extremely challenging. Here we describe a simple approach that allows determining the parent-specific methylation patterns of multiple genes in individual early embryos. Limiting dilution (LD) of bisulfite-treated DNA is combined with independent multiplex PCRs of single DNA target molecules to avoid amplification bias. Using this approach, we compared the methylation status of three imprinted (H19, Snrpn and Igf2r) and one pluripotency-related gene (Oct4) in three different groups of single mouse two-cell embryos. Standard in vitro fertilization of superovulated oocytes and the use of in vitro matured oocytes were not associated with significantly increased rates of stochastic single CpG methylation errors and epimutations (allele methylation errors), when compared with the in vivo produced controls. Similarly, we compared the methylation patterns of two imprinted genes (H19 and Snrpn) in individual mouse 16-cell embryos produced in vivo from superovulated and non-superovulated oocytes and did not observe major between-group differences. Using bovine oocytes and polar bodies as a model, we demonstrate that LD even allows the methylation analysis of multiple genes in single cells.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.