Nuclear Reprogramming: Methods and Protocols

ISBN-13: 9781493954414 / Angielski / Miękka / 2016 / 285 str.

Nathalie Beaujean; Helene Jammes; Alice Jouneau

Opis

Nuclear Reprogramming: Methods and Protocols, Second Edition includes not only classic methods to perform nuclear transfer in different species but also several techniques to assess the early and late development of the reconstructed embryos, at the cellular, molecular, and epigenetic level. Over the past several years, many technical improvements have been made to improve somatic cell nuclear transfer (SCNT) efficiency, all of which are reflected in the detailed chapters of this fully revised collection. 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.Authoritative and up-to-date, Nuclear Reprogramming: Methods and Protocols, Second Edition will be of interest not only to cloners but also to researchers concerned with studying the development of embryos.

Spis treści

1. Nuclear Transfer in the Mouse

            Vincent Brochard and Zichuan Liu

 

2. Nuclear Transfer in the Rabbit

            Nathalie Daniel and Patrick Chesné

 

3. Nuclear Transfer in Ruminants

            Joon-Hee Lee and Walid E. Maalouf

 

4. Nuclear Transfer and Transgenesis in the Pig

            Mayuko Kurome, Barbara Kessler, Annegret Wuensch, Hiroshi Nagashima, and Eckhard Wolf

 

5. Embryonic Stem Cell-Somatic Cell Fusion and Post-Fusion Enucleation

            Huseyin Sumer and Paul J. Verma

 

6. Analysis of Nuclear Reprogramming Following Nuclear Transfer to Xenopus Oocyte

            Jerome Jullien

 

7. Assessing the Quality of Donor Cells: Karyotyping Methods

            Amélie Bonnet-Garnier, Anne-Clemence Veillard, Bertrand Bed’Hom, Hélène Hayes, and Janice Britton-Davidian

 

8. Treatment of Donor Cell/Embryo with Different Approaches to Improve Development After Nuclear Transfer

            Eiji Mizutani, Sayaka Wakayama, and Teruhiko Wakayama

 

9. Fluorescent Immunodetection of Epigenetic Modifications on Pre-Implantation Mouse Embryos

            Claire Boulesteix and Nathalie Beaujean

 

10. Visualization of Epigenetic Modifications in Preimplantation Embryos

            Hiroshi Kimura and Kazuo Yamagata

 

11. Live Embryo Imaging to Follow Cell Cycle and Chromosomes Stability After Nuclear Transfer

            Sebastian T. Balbach and Michele Boiani

 

12. Analysis of Nucleolar Morphology and Protein Localization as an Indicator of Nuclear Reprogramming

            Olga Østrup, Hanne S. Pedersen, Hanne M. Holm, and Poul Hyttel

 

13. Assessment of Cell Lineages and Cell Death in Blastocysts by Immunostaining

            Claire Chazaud

 

14. Gene Expression Analysis in Early Embryos through Reverse Transcription Quantitative PCR (RT-qPCR)

            Nathalie Peynot, Véronique Duranthon, and Daulat Raheem Khan

 

15. Studying Bovine Early Embryo Transcriptome by Microarray

            Isabelle Dufort, Claude Robert, and Marc-André Sirard

 

16. Methylation of Specific Regions: Bisulfite-Sequencing at the Single Oocyte or 2-Cell Embryo Level

            Annick Lefèvre and Thierry Blachère

 

17. Micro Chromatin Immunoprecipitation (ChIP) from Early Mammalian Embryos

            John Arne Dahl and Arne Klungland

 

18. Assessing Reprogramming by Chimera Formation and Tetraploid Complementation

            Xin Li, Bao-long Xia, Wei Li, and Qi Zhou

 

19. Whole-Mount In Situ Hybridization to Assess Advancement of Development and Embryo Morphology

            Bernd Püschel and Alice Jouneau

 

20. Genome-Wide Analysis of Methylation in Bovine Clones by Methylated DNA Immunoprecipitation (MeDIP)

            Hélène Kiefer