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This volume provides an overview of zygotic genome activation (ZGA) and the use of recent tools to elucidate the events that occur during ZGA. The chapters in this book cover topics such as: clearance of maternal RNAs, detection of miRNA in mammalian oocytes and embryos, RNA FISH to study ZGA in early mouse embryos, and CRISPR/Cas9-mediated gene targeting during embryogenesis in the mouse and pig. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Thorough and cutting-edge, Zygotic Genome Activation: Methods and Protocols is a valuable resource for scientists and researchers who are interested in learning more about practical approaches to monitor the changes seen during ZGA.
List of contents
Clearance of Maternal RNAs: Not a Mummy's Embryo Anymore.- Link of Zygotic Genome Activation and Cell Cycle Control.- Role of MicroRNAs in Zygotic Genome Activation: Modulation of mRNA during Embryogenesis.- Gene Expression Analysis of Mammalian Oocytes and Embryos by Quantitative Real-Time RT-PCR.- Detection of miRNA in Mammalian Oocytes and Embryos.- Detection of Bidirectional Promoter-Derived IncRNAs from Small-Scale Samples using Pre-Amplification-Free Directional RNA-Seq Method.- Detection and Characterization of Small Non-Coding RNAs in Mouse Gametes and Embryos Prior to Zygotic Genome Activation.- Purification of Zygotically Transcribed RNA through Metabolic Labeling of Early Zebrafish Embryos.- RNA FISH to Study Zygotic Genome Activation in Early Mouse Embryos.- Detection of RNA Polymerase II in Mice Embryos during Zygotic Genome Activation using Immunocytochemistry.- Immunological Staining of Global Changes in DNA Methylation in the Early Mammalian Embryo.- Single Cell Restriction Enzyme-Based Analysis of Methylation at Genomic Imprinted Regions in Preimplantation Mouse Embryos.- Use of Chemicals to Inhibit DNA Replication, Transcription, and Protein Synthesis to Study Zygotic Genome Activation.- Targeted Gene Knockdown in Early Embryos using siRNA.- Generating Mouse Models using Zygote Electroporation of Nucleases (ZEN) Technology with High Efficiency and Throughput.- CRISPR/Cas9-Mediated Gene Targeting during Embryogenesis in Swine.- Potential Involvement of SCF-Complex in Zygotic Genome Activation during Early Bovine Embryo Development.- Use of Histone K-M Mutants for the Analysis of Transcriptional Regulation in Mouse Zygotes.
Summary
This volume provides an overview of zygotic genome activation (ZGA) and the use of recent tools to elucidate the events that occur during ZGA. The chapters in this book cover topics such as: clearance of maternal RNAs, detection of miRNA in mammalian oocytes and embryos, RNA FISH to study ZGA in early mouse embryos, and CRISPR/Cas9-mediated gene targeting during embryogenesis in the mouse and pig. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Thorough and cutting-edge, Zygotic Genome Activation: Methods and Protocols is a valuable resource for scientists and researchers who are interested in learning more about practical approaches to monitor the changes seen during ZGA.
