Nuclear Transfer Protocols


Book Description

Nuclear Transfer Protocols: Cell Reprogramming and Transgenesis is a comprehensive review of nuclear transfer technology in vertebrates, aimed at reprogramming differentiated nuclei and effecting targeted gene transfer. The emphasis here is on providing readily reproducible techniques for the gene- tion of cloned embryos and animals in a number of key research and commercially important vertebrates. Additional chapters provide alternative cutting-edge methods for nuclear transfer, such as zona-free nuclear transfer and serial nuclear transfer. Of immense practical benefit are descriptions of procedures associated with cloning, such as in vitro maturation of oocytes, activation and culture of cloned embryos, maintenance of pregnancy, and neonatal care of clones. Nuclear Transfer Protocols: Cell Reprogramming and Transgenesis also provides an understanding of the factors involved in nuclear reprogramming, which is imperative for the success of cloning. A section dealing with such cloning-related issues as aging and normality of clones is also included making this an essential comprehensive handbook for research and commercial labo- tories involved in, or intending to work on, nuclear transfer. The volume will prove beneficial to molecular biologists, stem cell biologists, clinicians, biotechnologists, students, veterinarians, and animal care technicians involved with reprogramming, nuclear transfer, and transgenesis.




Nuclear Transfer Protocols


Book Description

"Nuclear Transfer Protocols: Cell Reprogramming and Transgenesis" is a comprehensive review of nuclear transfer technology in vertebrates, aimed at reprogramming differentiated nuclei and effecting targeted gene transfer. The emphasis here is on providing readily reproducible techniques for the generation of cloned embryos and animals in a number of key research and commercially important vertebrates. Additional chapters provide alternative cutting-edge methods for nuclear transfer, such as zona-free nuclear transfer and post-fusion enucleation nuclear transfer.




Nuclear Reprogramming


Book Description

This volume provides basic and advanced protocols on somatic cell nuclear transfer, induced pluripotent stem cells, and direct reprogramming of somatic cells into different functional cells. Chapters guide readers through methods on standardized procedures for characterization of induced pluripotent stem cells, as well as those for preparation of materials required for induction of pluripotent stem cells. 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 cutting-edge, Nuclear Reprogramming: Methods and Protocols aims to ensure successful results in the further study of this vital field. .




Nuclear Reprogramming


Book Description

A wide-ranging collection of readily reproducible methods for performing nuclear reprogramming by nuclear transfer in several different species, by fusion through both chemical treatment and electrically shocking cells, and by in vivo treatment of cells with cell extracts. Several methods of monitoring nuclear reprogramming are also 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 verification of surface marker expression and the 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.




Somatic Cell Nuclear Transfer Technology


Book Description

This volume details protocols on animal cloning by Somatic cell nuclear transfer for basic research and biotechnological applications. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Somatic Cell Nuclear Transfer Technology aims to be comprehensive guide for researchers.




Nuclear Transfer Protocols


Book Description

Nuclear Transfer Protocols: Cell Reprogramming and Transgenesis is a comprehensive review of nuclear transfer technology in vertebrates, aimed at reprogramming differentiated nuclei and effecting targeted gene transfer. The emphasis here is on providing readily reproducible techniques for the gene- tion of cloned embryos and animals in a number of key research and commercially important vertebrates. Additional chapters provide alternative cutting-edge methods for nuclear transfer, such as zona-free nuclear transfer and serial nuclear transfer. Of immense practical benefit are descriptions of procedures associated with cloning, such as in vitro maturation of oocytes, activation and culture of cloned embryos, maintenance of pregnancy, and neonatal care of clones. Nuclear Transfer Protocols: Cell Reprogramming and Transgenesis also provides an understanding of the factors involved in nuclear reprogramming, which is imperative for the success of cloning. A section dealing with such cloning-related issues as aging and normality of clones is also included making this an essential comprehensive handbook for research and commercial labo- tories involved in, or intending to work on, nuclear transfer. The volume will prove beneficial to molecular biologists, stem cell biologists, clinicians, biotechnologists, students, veterinarians, and animal care technicians involved with reprogramming, nuclear transfer, and transgenesis.




Principles of Cloning


Book Description

Principles of Cloning, Second Edition is the fully revised edition of the authoritative book on the science of cloning. The book presents the basic biological mechanisms of how cloning works and progresses to discuss current and potential applications in basic biology, agriculture, biotechnology, and medicine. Beginning with the history and theory behind cloning, the book goes on to examine methods of micromanipulation, nuclear transfer, genetic modification, and pregnancy and neonatal care of cloned animals. The cloning of various species—including mice, sheep, cattle, and non-mammals—is considered as well. The Editors have been involved in a number of breakthroughs using cloning technique, including the first demonstration that cloning works in differentiated cells done by the Recipient of the 2012 Nobel Prize for Physiology or Medicine – Dr John Gurdon; the cloning of the first mammal from a somatic cell – Drs Keith Campbell and Ian Wilmut; the demonstration that cloning can reset the biological clock - Drs Michael West and Robert Lanza; the demonstration that a terminally differentiated cell can give rise to a whole new individual – Dr Rudolf Jaenisch and the cloning of the first transgenic bovine from a differentiated cell – Dr Jose Cibelli. The majority of the contributing authors are the principal investigators on each of the animal species cloned to date and are expertly qualified to present the state-of-the-art information in their respective areas. - First and most comprehensive book on animal cloning, 100% revised - Describes an in-depth analysis of current limitations of the technology and research areas to explore - Offers cloning applications on basic biology, agriculture, biotechnology, and medicine




Mitochondrial Replacement Techniques


Book Description

Mitochondrial replacement techniques (MRTs) are designed to prevent the transmission of mitochondrial DNA (mtDNA) diseases from mother to child. While MRTs, if effective, could satisfy a desire of women seeking to have a genetically related child without the risk of passing on mtDNA disease, the technique raises significant ethical and social issues. It would create offspring who have genetic material from two women, something never sanctioned in humans, and would create mitochondrial changes that could be heritable (in female offspring), and therefore passed on in perpetuity. The manipulation would be performed on eggs or embryos, would affect every cell of the resulting individual, and once carried out this genetic manipulation is not reversible. Mitochondrial Replacement Techniques considers the implications of manipulating mitochondrial content both in children born to women as a result of participating in these studies and in descendants of any female offspring. This study examines the ethical and social issues related to MRTs, outlines principles that would provide a framework and foundation for oversight of MRTs, and develops recommendations to inform the Food and Drug Administration's consideration of investigational new drug applications.




Nuclear Reprogramming


Book Description

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.




Bovine Reproduction


Book Description

Bovine Reproduction is a comprehensive, current reference providing information on all aspects of reproduction in the bull and cow. Offering fundamental knowledge on evaluating and restoring fertility in the bovine patient, the book also places information in the context of herd health where appropriate for a truly global view of bovine theriogenology. Printed in full color throughout, the book includes 83 chapters and more than 550 images, making it the most exhaustive reference available on this topic. Each section covers anatomy and physiology, breeding management, and reproductive surgery, as well as obstetrics and pregnancy wastage in the cow. Bovine Reproduction is a welcome resource for bovine practitioners, theriogenologists, and animal scientists, as well as veterinary students and residents with an interest in the cow.