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Improving Bovine Cloning Efficiency by Enhancing Reprogramming during Embryonic Genome Activation (EGA)

通过增强胚胎基因组激活 (EGA) 过程中的重编程来提高牛克隆效率

基本信息

批准号：
9923462
负责人：
Edward John Grow
金额：
$ 7.14万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-01 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9923462
关键词：
Address Agriculture Animal Model Assisted Reproductive Technology Award Bioinformatics Biological Assay Birth Blastocyst Transfer Cattle Cell Lineage Cell Nucleus Cell model Cells Chromatin Cloning Complement Complementary DNA DNA Sequence Data Data Set Defect Development Developmental Process Dissection Elements Embryo Embryo Cloning Embryo Transfer Embryology Embryonic Development Enhancers Environmental Impact Epigenetic Process Event Exhibits Failure Fertilization Fibrinogen Fibroblasts Gene Activation Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Enhancer Element Genetic Transcription Genome Genomics Germ Cells Goals Homeobox Human In Vitro Inner Cell Mass Knowledge Logic Mammals Maps Mediating Memory Messenger RNA Methods Microinjections Microscopy Modeling Monitor Mus Nature Nucleic Acid Regulatory Sequences Oocytes Phenotype Pregnancy Process Publishing Regulation Regulator Genes Reporter Reporting Research Residual state Resistance Rest Sheep Somatic Cell System Techniques Testing Time Totipotency Transcriptional Activation Work animal cloning base blastocyst combinatorial egg embryo culture embryo quality epigenomics fascinate genome-wide human model improved in vivo innovation mature animal practical application preimplantation preservation programs promoter reconstitution skills somatic cell nuclear transfer sperm cell technology development transcription factor zygote

项目摘要

This F32 proposal outlines research objectives to investigate the early gene expression regulation of bovine somatic cell nuclear transfer (SCNT) cloned embryos, and improve their developmental potential by harnessing our knowledge of IVF bovine preimplantation development. Based on several published reports, including my own work, my hypothesis is that failed embryonic genome activation (EGA) is the rate-limiting step negatively impacting the robust development of cloned animals. Because many EGA activated genes are involved in epigenetic reprogramming embryonic chromatin, failure to appropriately turn on these genes leads to developmental dysregulation and failure, often manifested much later in gestation. Building upon both our published work, showing that DUX transcription factors (TF) are the master regulators of gene expression at the egg-to-embryo transition, and utilizing my extensive unpublished preliminary data laying the ground work for our hypothesis in the bovine system, I will first address this problem using cutting-edge genomic, epigenomic, and embryological methods. First, I will test whether the putative enhancer regions I identified in IVF bovine embryos show stage-specific transcriptional activity in vivo. I will also dissect which TFs the putative enhancer regions are responsive to using a cellular system. Then I will identify open chromatin regions in bovine SCNT embryos and compare them to the high-quality IVF embryo open chromatin maps I have previously generated. This will help me understand how much of the SCNT developmental phenotype is caused by failure to activate EGA chromatin or failure to decommission somatic cell open chromatin. Finally, I will use our data that cow DUXC is a major activator of bovine EGA to test our hypothesis that DUXC deficiency can be overcome by ectopically expressing DUXC in SCNT embryos and that this will increase the efficiency of SCNT. With significance for human assisted reproductive technologies, development of large animal models of human embryogenesis, agricultural significance of cloning, and ecological impacts of endangered species preservation through SCNT, this work integrates innovative strategies to impact multiple fields.

本F32提案概述了研究早期基因表达调控的研究目标