Primary and Stem Cells

This book describes basic cell engineering methods, emphasizing stem cell applications, and use of the genetically modified stem cells in cell therapy and drug discovery. Together, the chapters introduce and offer insights on new techniques for engineering of stem cells and the delivery of transgenes into stem cells via various viral and non-viral systems. The book offers a guide to the types of manipulations currently available to create genetically engineered stem cells that suit any investigator's purpose, whether it's basic science investigation, creation of disease models and screens, or cells for therapeutic applications.

This book describes basic cell engineering methods, emphasizing stem cell applications, and use of the genetically modified stem cells in cell therapy and drug discovery. Together, the chapters introduce and offer insights on new techniques for engineering of stem cells and the delivery of transgenes into stem cells via various viral and non-viral systems. The book offers a guide to the types of manipulations currently available to create genetically engineered stem cells that suit any investigator's purpose, whether it's basic science investigation, creation of disease models and screens, or cells for therapeutic applications.

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 EllerstrEURom, Peter Sartipy, Raimund Strehl, and Johan Hyllner13. Screening and Drug Discovery 251Elizabeth A. Davenport and Robert S. AmesIndex 273