Primary and Stem Cells

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.

Foreword Michele Calos vii

Preface ix

Contributors xi

Part I: Cloning and Gene Delivery 1

1. DNA Assembly Technologies Based on Homologous Recombination 3
Billyana Tsvetanova, Lansha Peng, Xiquan Liang, Ke Li, Jian-Ping Yang, Tony Ho, Josh Shirley, Liewei Xu, Jason Potter, Wieslaw Kudlicki, Todd Peterson, and Federico Katzen

2. Multigene Assembly for Construction of Synthetic Operons: Creation and Delivery of an Optimized All-IN-One Expression Construct for Generating Mouse iPS Cells 19
Takefumi Sone, Yoko Takata, Hiroe Kishine, Taichi Andoh, Fumio Imamoto, and Jonathan D. Chesnut

3. Strategies for the Delivery of Naked DNA 37
Jennifer C. Moore and Ronald P. Hart

Part II: Nonintegrating Technologies 49

4. Episomal Vectors 51
Junying Yu

5. Nonintegrating DNA Virus 85
Chris Kemp and David Kuninger

6. Nonintegrating RNA Viruses 103
Noemi Fusaki

7. Protein Delivery 119
Xiao-Jian Han and Kazuhito Tomizawa

Part III: Integrating Technologies 135

8. Sleeping Beauty Transposon-Mediated Stable Gene Delivery 137
Andrew Wilber, Xianzheng Zhou, Xin Huang, Dan S. Kaufman, and R. Scott McIvor

9. Integrating Viral Vectors for Gene Modifications 163
Gemma L. Mendel, Brian Paszkiet, and Rafal P. Witek

10. Bacteriophage Integrases for Site-Specific Integration 199
Bhaskar Thyagarajan

11. Improving Gene Targeting Efficiency in Human Pluripotent Stem Cells 211
Sangyoon Han, Anna McCann, Louise C. Laurent, Jeanne F. Loring, and Ying Liu

Part IV: Applications 227

12. Modified Stem Cells as Disease Models and in Toxicology Screening 229
Catharina Ellerstr€om, Peter Sartipy, Raimund Strehl, and Johan Hyllner

13. Screening and Drug Discovery 251
Elizabeth A. Davenport and Robert S. Ames

Index 273