Contents Preface Contributors Biallelic Gene Knockouts in Chinese Hamster Ovary Cells Naoko Yamane-Ohnuki, Kazuya Yamano, and Mitsuo Satoh Highly Proficient Gene Targeting by Homologous Recombination in the Human pre-B Cell Line Nalm-6 Noritaka Adachi, Aya Kurosawa, and Hideki Koyama Robust Cell Line Development Using Meganucleases Jean-Pierre Cabaniols and Frédéric Pâques Design and Testing of Zinc Finger Nucleases for Use in Mammalian Cells Matthew Porteus Gene Targeting in Drosophila and C. elegans with Zinc-finger Nucleases Dana Carroll, Kelly J. Beumer, J. Jason Morton, Ana Bozas and Jonathan K. Trautman Orpheus Recombination: A Comprehensive Bacteriophage System for Murine Targeting Vector Construction by Transplacement Knut Woltjen, Kenichi Ito, Teruhisa Tsuzuki, and Derrick E. Rancourt Transposon Mediated Mutagenesis in Somatic Cells: Identification of transposon-genomic DNA junctions David A. Largaespada and Lara S. Collier Insertional Mutagenesis of the Mouse Germline with Sleeping Beauty Transposition Junji Takeda, Zsuzsanna Izsvák, and Zoltán Ivics Conditional Gene Trapping Using the FLEx System Thomas Floss and Frank Schnütgen Steps Towards Targeted Insertional Mutagenesis with Class II Transposable Elements Sareina Chiung-Yuan Wu, Kommineni J. Maragathavally, Craig J. Coates and Joseph M. Kaminski Targeting integration of the Saccharomyces Ty5 retrotransposon Troy L. Brady, Clarice L. Schmidt and Daniel F. Voytas Site-Specific Chromosomal Integration Mediated by f C31 Integrase Annahita Keravala and Michele P. Calos Triplex-Mediated Gene Modification Erica B. Schleifman, Joanna Y. Chin, and Peter M.Glazer Allelic exchange of unmarked mutations in Mycobacterium tuberculosis. Martin S. Pavelka, Jr. Mycobacterial Recombineering Julia C. van Kessel and Graham F. Hatfull Chromosomal Engineering of Clostridium perfringens Using Group II Introns Phalguni Gupta and Yue Chen
Great disparities exist between organisms with regard to the relative ease of chromosomal mutagenesis and manipulation. In Chromosomal Mutagenesis, a team of experts provide a variety of chromosomal manipulation techniques, including insertional gene disruptions, gene knockouts, stimulated homologous recombination techniques and other novel tools, for both prokaryotic and eukaryotic organisms, and attempt to expand the genetic toolbox beyond model organisms. Following the format of the highly successful Methods in Molecular Biology(TM) format, each chapter offers step-by-step laboratory instructions, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and cutting-edge, Chromosomal Mutagenesis covers state-of-the-art techniques that are staged to expand, if not revolutionize, genetic analysis in the long neglected and relevant cell types.
Highlights techniques involving a wider variety of cell types and organisms
Contains state-of-the-art methods in step-by-step laboratory format