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Genome Editing and Engineering
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Table of Contents

List of contributors; Forewords; Preface; List of abbreviations; Part I. Biology of Endonucleases (Zinc-Finger Nuclease, TALENs and CRISPRs) and Regulatory Networks: 1. Introduction to genome editing and engineering: from Talens, ZFNs and CRISPRs to molecular surgery; 2. Targeted genome editing techniques in C. elegans and other nematode species; 3. Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors; 4. In vivo studies of miRNA target interactions using site-specific genome engineering; 5. Don't kill the messenger: employing genome editing to study regulatory RNA interactions; Part II. Genome Editing in Model Organisms: 6. Genome editing in the crustacean Daphnia magna using CRISPR/Cas and TALEN systems; 7. Leapfrogging: a method for targeting genome editing to the germline; 8. Genome editing with desired mutations (knock-in) by CRISPR in model organisms; 9. Genetically engineered pig models for human diseases using ZFNs, TALENs and CRISPR-Cas9; 10. Gene editing to create agricultural and biomedical swine models; 11. Generation of new model cell lines using ssODN knock-in donors and FACS-based genome editing; Part III. Technology Development and Screening: 12. CRISPR genome editing in mice; 13. Detection of insertion/deletion (indel) events after genome targeting: pros and cons of the available methods; 14. Application of TAL proteins and the CRISPR system to purification of specific genomic regions for locus-specific identification of chromatin-associated molecules; 15. Application of CRISPR for pooled, vector-based functional genomic screening in mammalian cell lines; 16. Generation and utilization of CRISPR/Cas9 screening libraries in mammalian cells; Part IV. Genome Editing in Stem Cells and Regenerative Biology: 17. Targeted genome editing using nuclease assisted vector integration; 18. Genome engineering using Sleeping Beauty transposition in vertebrates; 19. Genome editing of pluripotent stem cells; 20. Principles for targeting adult tissue stem cells to achieve durable gene and gene-editing therapies; 21. Therapeutic genome editing in human hematopoietic stem and progenitor cells; Part V. Genome Editing in Disease Biology: 22. CRISPR/Cas9-Based in vivo models of cancer; 23. Inducible CRISPR-based genome editing for the characterization of cancer genes; 24. Genome editing for retinal diseases; 25. Manipulation of long non-coding RNAs in cardiovascular disease using genome-editing technology; 26. Gene silencing, disruption and latency reactivation with RNA-based and gene editing CRISPR/Cas, ZFN and TALEN technologies for HIV-1/AIDS therapies; 27. Use of the CRISPR-Cas9 system for genome editing of immune system cells, defense against HIV-1 and cancer therapies; 28. Harnessing therapeutic potential of long non-coding RNAs in immunity; Part VI. Legal (Intelluctual Property) and Bioethical Issues of Genome Editing: 29. Patening of CRISPR-based inventions; 30. Patent, ownership, and licensing issues of CRISPR-based genome editing: impact on universities and their licensees; 31. The ethics of human genome editing; 32. Regulating human genome editing: negotiating ethical concerns through science and policy; Index.

Promotional Information

A complete guide to endonuclease-based genomic engineering, from basic science to application in disease biology and clinical treatment.

About the Author

Krishnarao Appasani is the Founder and Chief Executive Officer of GeneExpression Systems. He is an award winning scientist, and has edited several books, including: Optogenetics: From neuronal function to mapping and disease biology (Cambridge, 2017); Genome-Wide Association Studies: From Polymorphism to Personalized Medicine (Cambridge, 2016); Epigenomics: From Chromatin Biology to Therapeutics (Cambridge, 2012); MicroRNAs: From Basic Science to Disease Biology (Cambridge, 2007); and RNA Interference: From Basic Science to Drug Development (Cambridge, 2005).

Reviews

'This comprehensive book is welcome at this point of rapid growth in new technologies and discoveries such that keeping up with applications of genome engineering is challenging. Here we have both a general introduction to the field and to deep mechanisms and glimpses of future applications and ethical considerations - with gems for newcomers and experts alike.' George M. Church, Harvard University, Massachusetts and Massachusetts Institute of Technology
'Krishnarao Appasani has assembled a comprehensive series of up-to-date chapters on endonuclease-based genomic engineering by experts and practitioners. The book presents a wide range of topics from basic science, applications to genetic screens and diseases, and legal and ethical issues. The many potential applications across a wide range of organisms are well illustrated. The book will become a must have for any researchers in academia and industry trying to keep up with this fast-moving field.' Norbert Perrimon, Harvard Medical School, Boston
'... a remarkable book, which very successfully manages to provide deep insights into the various and latest aspects of gene editing and engineering. It is so far probably the best gene editing text collection available and highly suited to get a good understanding of the breadth and impact of this cutting-edge field of science.' Rodger Novak, President, Director and Chair of the Board CRISPR Therapeutics AG, Switzerland
'... offers a widespread analysis of the state of the art concerning all the main tools available for modification of eukaryotic DNA in vivo, their application in metazoans and the societal issues involved. I thoroughly recommend it to anyone interested in gaining insight into this newest genetic revolution.' Francisco Juan Martinez Mojica, Universidad de Alicante, Spain
'The field of precise genome engineering has been compiled into an excellent and essential guide through its landscape and it imparts state-of-the-art knowledge of its tools, its multiple research and clinical applications and related legal and ethical implications.' Shamim Herbert Rahman, Senior Research Scientist, Novo Nordisk A/S, Malov, Denmark
'... broadly covers recent development of genome editing from principle, history and usage to patent wars.' Keiji Nishida, Kobe University, Japan
'... providing significant and useful information on current knowledge in the field and promoting the interaction between basic and pre-clinical/clinical research ... of potential interest to a broad readership interested in basic and translational research.' Antonio Russo, 'P. Giaccone' University Hospital, University of Palermo, Italy
'This comprehensive book is welcome at this point of rapid growth in new technologies and discoveries such that keeping up with applications of genome engineering is challenging. Here we have both a general introduction to the field and to deep mechanisms and glimpses of future applications and ethical considerations - with gems for newcomers and experts alike.' George M. Church, Harvard University, Massachusetts and Massachusetts Institute of Technology
'Krishnarao Appasani has assembled a comprehensive series of up-to-date chapters on endonuclease-based genomic engineering by experts and practitioners. The book presents a wide range of topics from basic science, applications to genetic screens and diseases, and legal and ethical issues. The many potential applications across a wide range of organisms are well illustrated. The book will become a must have for any researchers in academia and industry trying to keep up with this fast-moving field.' Norbert Perrimon, Harvard Medical School, Boston
'... a remarkable book, which very successfully manages to provide deep insights into the various and latest aspects of gene editing and engineering. It is so far probably the best gene editing text collection available and highly suited to get a good understanding of the breadth and impact of this cutting-edge field of science.' Rodger Novak, President, Director and Chair of the Board CRISPR Therapeutics AG, Switzerland
'... offers a widespread analysis of the state of the art concerning all the main tools available for modification of eukaryotic DNA in vivo, their application in metazoans and the societal issues involved. I thoroughly recommend it to anyone interested in gaining insight into this newest genetic revolution.' Francisco Juan Martinez Mojica, Universidad de Alicante, Spain
'The field of precise genome engineering has been compiled into an excellent and essential guide through its landscape and it imparts state-of-the-art knowledge of its tools, its multiple research and clinical applications and related legal and ethical implications.' Shamim Herbert Rahman, Senior Research Scientist, Novo Nordisk A/S, Malov, Denmark
'... broadly covers recent development of genome editing from principle, history and usage to patent wars.' Keiji Nishida, Kobe University, Japan
'... providing significant and useful information on current knowledge in the field and promoting the interaction between basic and pre-clinical/clinical research ... of potential interest to a broad readership interested in basic and translational research.' Antonio Russo, 'P. Giaccone' University Hospital, University of Palermo, Italy

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