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Nuclear Reprogramming

Methods and Protocols

  • Steve Pells

Part of the Methods in Molecular Biology™ book series (MIMB, volume 325)

Table of contents

  1. Front Matter
    Pages i-xiv
  2. John B. Gurdon
    Pages 1-9
  3. William A. Ritchie
    Pages 11-23
  4. Shaorong Gao
    Pages 25-33
  5. Andrew C. Boquest, Aboulghassem Shahdadfar, Jan E. Brinchmann, Philippe Collas
    Pages 35-46
  6. Robert H. Broyles, Austin C. Roth, Mairead Todd, Visar Belegu
    Pages 47-57
  7. Jian Yang, Ming Hong Shen
    Pages 59-66
  8. Stephen Sullivan, Martin Waterfall, Ed J. Gallagher, Jim McWhir, Steve Pells
    Pages 81-97
  9. Anne-Mari Håkelien, Kristine G. Gaustad, Philippe Collas
    Pages 99-114
  10. Megan Munsie, Peter Mountford, Jennifer Nichols
    Pages 115-128
  11. Dean H. Betts, Steven Perrault, Lea Harrington, W. Allan King
    Pages 149-180
  12. Cornelia Wiese, Gabriela Kania, Alexandra Rolletschek, Przemyslaw Blyszczuk, Anna M. Wobus
    Pages 181-205
  13. Bruce A. Fenderson, Maria P. De Miguel, April D. Pyle, Peter J. Donovan
    Pages 207-224
  14. Hannah R. Moore, Richard R. Meehan, Lorraine E. Young
    Pages 239-249
  15. Fiona B. Turner, Wang L. Cheung, Peter Cheung
    Pages 261-272
  16. Hiromi Tagoh, Peter N. Cockerill, Constanze Bonifer
    Pages 285-314
  17. Back Matter
    Pages 327-333

About this book

Introduction

Although nuclear reprogramming-the cellular process underlying cloning and the artificial generation of stem cells from somatic cells-may be considered a "black box" whose interior events are either completely unknown or at least imperfectly characterized, tantalizing clues about its operation are beginning to emerge. In Nuclear Reprogramming: Methods and Protocols, hands-on laboratory scientists describe readily reproducible methods for actually performing nuclear reprogramming by nuclear transfer in several different species, by fusion induced both chemically and electrically, and by the in vivo treatment of cells with cell extracts. Isolation of an adult stem cell type is also described. In addition, several methods of monitoring nuclear reprogramming are presented, including the use of transgenic markers, activation of telomerase as an ES-specific marker, light and electron microscopic observation of structural changes in the nucleus, and the verification of surface marker expression and differentiation potential of stem cells. Biochemical methods are provided for the examination of chromatin protein modifications, nucleosomal footprinting, transcription factor binding, and the study of DNA methylation changes both at the specific locus level and at the level of the whole nucleus. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Wide-ranging and highly practical, Nuclear Reprogramming: Methods and Protocols significantly expands the repertoire of methods available to perform, study, and understand nuclear programming, not only for cloning, but also for reprogramming somatic cells to stem cells.

Keywords

Activation DNA Ethylen Monoclonal Antibodies PCR Telomere Vivo transcription

Editors and affiliations

  • Steve Pells
    • 1
  1. 1.Department of Gene Function and DevelopmentRoslin InstituteRoslin

Bibliographic information

  • DOI https://doi.org/10.1385/1597450057
  • Copyright Information Humana Press 2006
  • Publisher Name Humana Press
  • eBook Packages Springer Protocols
  • Print ISBN 978-1-58829-379-4
  • Online ISBN 978-1-59745-005-8
  • Series Print ISSN 1064-3745
  • Series Online ISSN 1940-6029
  • Buy this book on publisher's site