Gene Transfer Methods: Introducing DNA Into Living Cells and Organisms 

1-881299-34-1

978-1-881299-34-9

P.A. Norton and L.F. Steel, ed., Eaton Publishing, 2000, 272 pp., hard cover

(partial)
1. AN INTRODUCTION TO GENE TRANSFER
2.TRANSFORMATION OF BACTERIA AND PURIFICATION OF PLASMID
3. YEAST TRANSFORMATION
4. TRANSFORMATION OF DICTYOSTELIUM
II. Transformation Techniques
III. Copy Number and Vector Integration Site
IV. Drug and Nutritional Selections
V. Applications
VI. Protocols
VII. Stock Solutions
VIII. Strains and Vectors
5. INTRODUCTION OF DNA INTO CULTURE MAMMALIAN CELLS
III. Mechanisms of DNA Uptake
VI. Procedures
A. Protocol for DEAE Dextran-Mediated Transfection
B. Protocol for CaPO4-Mediated Transfection
C. Protocol for Lipid-Mediated DNA Uptake
D. Protocol for PEI-Mediated DNA Uptake
E. Protocol for Electroporation
F. Measuring Transfection Efficiency by X-Gal Staining
G. Protocol for Stable Transfections Using G418 Selection
6. GENERATION OF TRANSGENIC DROSOPHILA MELANOGASTER BY P ELEMENT-MEDIATED GERMLINE TRANSFORMATION
II. General Considerations
III. Protocols
IV. Expected Results and Troubleshooting
7. INTRODUCTION OF DNA INTO MOUSE EMBRYOS
A. Advantages of Exogenous Transgenes
B. Methods of Introducing Exogenous Transgenes into Embryos
C. Mechanisms of Exogenous Transgene Integration into Genomic DNA
III. Modifying Endogenous Genes
A. General Design of Gene Knockout Vectors
B. Pluripotent Embryonic Stem Cells
IV. Protocols
A. Growth and Maintenance of ES cells
B. Deriving ES Cells from Delayed Blastocysts
C. Transfection and Screening ES Cells
D. Introducing ES Cells into Embryos
E. Deriving Homozygous Mutant ES Cells via Loss of Heterozygosity: Implications for Sequential Gene Targeting Strategies
V. Conditional and Intra-Gene Targeting
A. Obtaining Intra-Gene Mutations with the Cre–LoxP Recombinase System
B. Obtaining Conditional Mutations by Breeding with Cre-Expressing Transgenic Mice
8. INTRODUCTION OF DNA INTO PLANTS
II. Gene Transfer Systems
A. Classical Agrobacterium System
B. In Planta Agrobacterium System
C. Direct Gene Transfer by Particle Bombardment
III. Protocols
A. Classical Agrobacterium System
B. In Planta Agrobacterium System for Arabidopsis
C. Particle Bombardment
IV. Media, Solutions, and Reagents
A. Classical Agrobacterium-Mediated Transformation
B. In Planta Agrobacterium-Mediated Transformation
9. INTRODUCTION OF THERAPEUTIC cDNAs INTO THE LUNGS OF HUMANS AND ANIMALS
II. Vectors for Lung Gene Transfer
A. Viral Vectors
B. Nonviral Vectors
III. In Vivo Methods of Administration
A. Ex Vivo Delivery
B. Intravenous Delivery
C. Direct Lung Instillation
D. Aerosol Administration
IV. Barriers to Airway Gene Transfer
A. Nonspecific Barriers to Gene Transfer
B. Vector-Specific Barriers to Luminal Airway Gene Transfer
C. Inflammatory and Immune Barriers to Airway Epithelial Gene Transfer
V. Strategies to Overcome Apical Membrane Barriers to Efficient Transduction
A. Host Modification
B. Vector Modification
C. Circumventing the Immune Response to Enhance Gene Transfer
10. GENETIC MANIPULATION AND THE PRECAUTIONARY PRINCIPLE
II. Three Hazardous Gene Modification Technologies
A. Microorganisms Containing Animal Genes
B. Genetically Modified Crops
C. Human Developmental Gene Modification
III. The Need for a Precautionary Science