UMA LAKSHMIPATHY is a principal investigator at Life Technologies. She has a PhD in life sciences, with academic and industry experience in molecular biology and stem cells. Dr. Lakshmipathy holds four patents and has authored more than forty publications. BHASKAR THYAGARAJAN is a program manager at Life Technologies. He has a PhD in pharmacology, with expertise in the areas of molecular biology, DNA recombination, gene and cell therapy, and protein purification. He holds one patent and has authored more than twenty publications.

“This is an excellent resource on the new techniques for the engineering of stem cells and the delivery of transgenes into stem cells via various viral and nonviral systems. It provides comprehensive information on the concepts of cell engineering principles and their application in primary and stem cells.  Researchers, established stem cell scientists, and graduate students interested in this field will find this book very handy.”  (Doody’s, 17 August 2012)

• Foreword Michele Calos viiPreface ixContributors xiPart I: Cloning and Gene Delivery 11. DNA Assembly Technologies Based on Homologous Recombination 3Billyana Tsvetanova, Lansha Peng, Xiquan Liang, Ke Li, Jian-Ping Yang, Tony Ho, Josh Shirley, Liewei Xu, Jason Potter, Wieslaw Kudlicki, Todd Peterson, and Federico Katzen2. Multigene Assembly for Construction of Synthetic Operons: Creation and Delivery of an Optimized All-IN-One Expression Construct for Generating Mouse iPS Cells 19Takefumi Sone, Yoko Takata, Hiroe Kishine, Taichi Andoh, Fumio Imamoto, and Jonathan D. Chesnut3. Strategies for the Delivery of Naked DNA 37Jennifer C. Moore and Ronald P. HartPart II: Nonintegrating Technologies 494. Episomal Vectors 51Junying Yu5. Nonintegrating DNA Virus 85Chris Kemp and David Kuninger6. Nonintegrating RNA Viruses 103Noemi Fusaki7. Protein Delivery 119Xiao-Jian Han and Kazuhito TomizawaPart III: Integrating Technologies 1358. Sleeping Beauty Transposon-Mediated Stable Gene Delivery 137Andrew Wilber, Xianzheng Zhou, Xin Huang, Dan S. Kaufman, and R. Scott McIvor9. Integrating Viral Vectors for Gene Modifications 163Gemma L. Mendel, Brian Paszkiet, and Rafal P. Witek10. Bacteriophage Integrases for Site-Specific Integration 199Bhaskar Thyagarajan11. Improving Gene Targeting Efficiency in Human Pluripotent Stem Cells 211Sangyoon Han, Anna McCann, Louise C. Laurent, Jeanne F. Loring, and Ying LiuPart IV: Applications 22712. Modified Stem Cells as Disease Models and in Toxicology Screening 229Catharina Ellerstr€om, Peter Sartipy, Raimund Strehl, and Johan Hyllner13. Screening and Drug Discovery 251Elizabeth A. Davenport and Robert S. AmesIndex 273

UMA LAKSHMIPATHY is a principal investigator at Life Technologies. She has a PhD in life sciences, with academic and industry experience in molecular biology and stem cells. Dr. Lakshmipathy holds four patents and has authored more than forty publications. BHASKAR THYAGARAJAN is a program manager at Life Technologies. He has a PhD in pharmacology, with expertise in the areas of molecular biology, DNA recombination, gene and cell therapy, and protein purification. He holds one patent and has authored more than twenty publications.

“This is an excellent resource on the new techniques for the engineering of stem cells and the delivery of transgenes into stem cells via various viral and nonviral systems. It provides comprehensive information on the concepts of cell engineering principles and their application in primary and stem cells.  Researchers, established stem cell scientists, and graduate students interested in this field will find this book very handy.”  (Doody’s, 17 August 2012)

• Foreword Michele Calos viiPreface ixContributors xiPart I: Cloning and Gene Delivery 11. DNA Assembly Technologies Based on Homologous Recombination 3Billyana Tsvetanova, Lansha Peng, Xiquan Liang, Ke Li, Jian-Ping Yang, Tony Ho, Josh Shirley, Liewei Xu, Jason Potter, Wieslaw Kudlicki, Todd Peterson, and Federico Katzen2. Multigene Assembly for Construction of Synthetic Operons: Creation and Delivery of an Optimized All-IN-One Expression Construct for Generating Mouse iPS Cells 19Takefumi Sone, Yoko Takata, Hiroe Kishine, Taichi Andoh, Fumio Imamoto, and Jonathan D. Chesnut3. Strategies for the Delivery of Naked DNA 37Jennifer C. Moore and Ronald P. HartPart II: Nonintegrating Technologies 494. Episomal Vectors 51Junying Yu5. Nonintegrating DNA Virus 85Chris Kemp and David Kuninger6. Nonintegrating RNA Viruses 103Noemi Fusaki7. Protein Delivery 119Xiao-Jian Han and Kazuhito TomizawaPart III: Integrating Technologies 1358. Sleeping Beauty Transposon-Mediated Stable Gene Delivery 137Andrew Wilber, Xianzheng Zhou, Xin Huang, Dan S. Kaufman, and R. Scott McIvor9. Integrating Viral Vectors for Gene Modifications 163Gemma L. Mendel, Brian Paszkiet, and Rafal P. Witek10. Bacteriophage Integrases for Site-Specific Integration 199Bhaskar Thyagarajan11. Improving Gene Targeting Efficiency in Human Pluripotent Stem Cells 211Sangyoon Han, Anna McCann, Louise C. Laurent, Jeanne F. Loring, and Ying LiuPart IV: Applications 22712. Modified Stem Cells as Disease Models and in Toxicology Screening 229Catharina Ellerstr€om, Peter Sartipy, Raimund Strehl, and Johan Hyllner13. Screening and Drug Discovery 251Elizabeth A. Davenport and Robert S. AmesIndex 273

UMA LAKSHMIPATHY is a principal investigator at Life Technologies. She has a PhD in life sciences, with academic and industry experience in molecular biology and stem cells. Dr. Lakshmipathy holds four patents and has authored more than forty publications. BHASKAR THYAGARAJAN is a program manager at Life Technologies. He has a PhD in pharmacology, with expertise in the areas of molecular biology, DNA recombination, gene and cell therapy, and protein purification. He holds one patent and has authored more than twenty publications.

“This is an excellent resource on the new techniques for the engineering of stem cells and the delivery of transgenes into stem cells via various viral and nonviral systems. It provides comprehensive information on the concepts of cell engineering principles and their application in primary and stem cells.  Researchers, established stem cell scientists, and graduate students interested in this field will find this book very handy.”  (Doody’s, 17 August 2012)

• Foreword Michele Calos viiPreface ixContributors xiPart I: Cloning and Gene Delivery 11. DNA Assembly Technologies Based on Homologous Recombination 3Billyana Tsvetanova, Lansha Peng, Xiquan Liang, Ke Li, Jian-Ping Yang, Tony Ho, Josh Shirley, Liewei Xu, Jason Potter, Wieslaw Kudlicki, Todd Peterson, and Federico Katzen2. Multigene Assembly for Construction of Synthetic Operons: Creation and Delivery of an Optimized All-IN-One Expression Construct for Generating Mouse iPS Cells 19Takefumi Sone, Yoko Takata, Hiroe Kishine, Taichi Andoh, Fumio Imamoto, and Jonathan D. Chesnut3. Strategies for the Delivery of Naked DNA 37Jennifer C. Moore and Ronald P. HartPart II: Nonintegrating Technologies 494. Episomal Vectors 51Junying Yu5. Nonintegrating DNA Virus 85Chris Kemp and David Kuninger6. Nonintegrating RNA Viruses 103Noemi Fusaki7. Protein Delivery 119Xiao-Jian Han and Kazuhito TomizawaPart III: Integrating Technologies 1358. Sleeping Beauty Transposon-Mediated Stable Gene Delivery 137Andrew Wilber, Xianzheng Zhou, Xin Huang, Dan S. Kaufman, and R. Scott McIvor9. Integrating Viral Vectors for Gene Modifications 163Gemma L. Mendel, Brian Paszkiet, and Rafal P. Witek10. Bacteriophage Integrases for Site-Specific Integration 199Bhaskar Thyagarajan11. Improving Gene Targeting Efficiency in Human Pluripotent Stem Cells 211Sangyoon Han, Anna McCann, Louise C. Laurent, Jeanne F. Loring, and Ying LiuPart IV: Applications 22712. Modified Stem Cells as Disease Models and in Toxicology Screening 229Catharina Ellerstr€om, Peter Sartipy, Raimund Strehl, and Johan Hyllner13. Screening and Drug Discovery 251Elizabeth A. Davenport and Robert S. AmesIndex 273