Primary and Stem Cells: Gene Transfer Technologies and Applications

The all–important resource that lays the groundwork forresearchers to genetically modify primary and stem cells for use indownstream applications

Replacing damaged or lost organs by producing "spare parts" is aconcept that lies at the core of the varied biotechnologicalpractices generally referred to as tissue engineering. An importantcomponent in this regeneration process includes the use of stemcells, which harbor the ability to mature into other cell types andhold the potential to act as a repair system for the body.Primary and Stem Cells describes basic cell engineeringmethods, emphasizing stem cell applications, and the use ofgenetically modified stem cells in cell therapy and drug discovery.Discussion includes new techniques for the engineering of stemcells and the delivery of transgenes into stem cells via variousviral and non–viral systems, along with:

Transfection techniques and methods for cell lineengineering

Information that assists researchers and drug developers indeciding how to generate their required drug or gene therapists indeveloping genetically modified cells

An overview of applications in areas like disease models anddrug discovery

Issues that address the interests of companies involved in stemcell tools and reagents

Primary and Stem Cells serves as an essential guide tothe types of manipulations currently available to creategenetically engineered stem cells that suit any investigator′spurpose basic science investigation, creation of diseasemodels and screens, or cells for therapeutic applications andacts as the launching pad for pioneering further development in afield emerging with exciting possibilities.

FOREWORD vii
Michele Calos

PREFACE ix

CONTRIBUTORS xi

PART I: CLONING AND GENE DELIVERY 1

1. DNA Assembly Technologies Based on Homologous Recombination3
Billyana Tsvetanova, Lansha Peng, Xiquan Liang, Ke Li, Jian–PingYang, Tony Ho, Josh Shirley, Liewei Xu, Jason Potter, WieslawKudlicki, Todd Peterson, and Federico Katzen

2. Multigene Assembly for Construction of Synthetic Operons:Creation and Delivery of an Optimized All–IN–One ExpressionConstruct for Generating Mouse iPS Cells 19
Takefumi Sone, Yoko Takata, Hiroe Kishine, Taichi Andoh, FumioImamoto, 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 GeneDelivery 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 Integration199
Bhaskar Thyagarajan

11. Improving Gene Targeting Efficiency in Human PluripotentStem 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 ToxicologyScreening 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

UMA LAKSHMIPATHY is a principal investigator at LifeTechnologies. She has a PhD in life sciences, with academic andindustry experience in molecular biology and stem cells. Dr.Lakshmipathy holds four patents and has authored more than fortypublications.

BHASKAR THYAGARAJAN is a program manager at LifeTechnologies. He has a PhD in pharmacology, with expertise in theareas of molecular biology, DNA recombination, gene and celltherapy, and protein purification. He holds one patent and hasauthored more than twenty publications.

This is an excellent resource on the new techniques forthe engineering of stem cells and the delivery of transgenes intostem cells via various viral and nonviral systems. It providescomprehensive information on the concepts of cell engineeringprinciples and their application in primary and stem cells. Researchers, established stem cell scientists, and graduatestudents interested in this field will find this book veryhandy.   (Doody s, 17 August 2012)