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Gene Expression in Recombinant Animal Cells and Transgenic Animals

  1. Richard M. Twyman1,
  2. Bruce Whitelaw2

Published Online: 15 APR 2010

DOI: 10.1002/9780470054581.eib342

Encyclopedia of Industrial Biotechnology

Encyclopedia of Industrial Biotechnology

How to Cite

M. Twyman, R. and Whitelaw, B. 2010. Gene Expression in Recombinant Animal Cells and Transgenic Animals. Encyclopedia of Industrial Biotechnology. 213–295.

Author Information

  1. 1

    John Inness Centre, Norwich, United Kingdom

  2. 2

    Roslin Institute, Roslin, Midlothian, United Kingdom

Publication History

  1. Published Online: 15 APR 2010

Abstract

  • Introduction

  • Overview of Animal Cell Technology

    • Animal Cells Used for Genetic Manipulation

    • Protein Expression

    • Recent Advances in Gene Transfer Technology

    • Transfer and Fate of DNA Introduced into Animal Cells

  • Plasmid Expression Vectors for Animal Cells

    • Classes of Expression Vector and Common Modular Components

    • Plasmid Backbone Sequences

    • Plasmid Maintenance Sequences

    • Regulatory Sequences

    • Marker Genes

    • Sequences to Simplify Protein Purification

  • Other Direct Transfer Vectors for Animal Cells

    • Yeast Artificial Chromosome Expression Vectors

    • Targeting Vectors

    • Plasmid Vectors for Insertional Mutagenesis and Entrapment

  • Direct DNA Transfer to Animal Cells

    • Overview of Animal Cell Transfection

    • Transient Transfection with Nonreplicating Vectors

    • Transient Transfection with Runaway Polyomavirus Replicons

    • Reporter Genes in Transfection Analysis

    • Stable Transfection Using Nonreplicating DNA

    • Selectable Markers for Use in Mammalian Cell Lines

    • Maximizing Transgene Expression by Gene Amplification

    • Stable Transfection Using Episomally Maintained Vectors

  • Transfection Methods

    • Overview of Transfection Strategies

    • Transfection with Locally Applied Naked DNA

    • Transfection with Calcium Phosphate

    • Transfection with DEAE–Dextran

    • Enhancing Transient Transfection Efficiency

    • Electroporation and Laser Transfection

    • Liposomes, Virosomes, and Lipofection

    • Polybrene-Mediated Transfection

    • Cell or Protoplast Fusion

    • Microinjection

    • Transfection by Particle Bombardment

    • Receptor-Mediated Transfection

  • Viral Expression Vectors for Animal Cells

    • General Properties of Viral Vectors

    • Adenovirus Vectors

    • Adeno-Associated Virus Vectors

    • Alphavirus Vectors

    • Baculovirus Vectors

    • Herpes Virus Vectors

    • Retrovirus Vectors

    • SV40 Vectors

    • Vaccinia and Other Poxvirus Vectors

    • Hybrid Viral Vectors

  • Expression Parameters and Optimization—Vector and Insert Sequences

    • Transcriptional Termination and RNA Processing

    • Protein Synthesis

    • Post-Translational Events

  • Expression Parameters and Optimization—Consequences of Transgene Integration

    • Overview

    • Timing of Integration

    • The Fate of Foreign DNA and the Structure of the Transgenome

    • Position Effects

    • Integration Effects

    • Transgene Copy Number and Dosage Effects

    • Transgene Organization

    • DNA Methylation and Homology-Dependent Gene Silencing

    • Transgene Stability and Transmission

    • Optimal Design of Transgenes for Use in Transgenic Animals

    • Overcoming Position Effects

  • Recombination-Based Methodology and Gene Inhibition Strategy

    • Recombination and Its Use in Genetic Engineering

    • Strategies for the Inhibition of Endogenous Genes

    • Homologous Recombination

    • Early Development of Homologous Recombination as a Genetic Tool

    • Gene Silencing—Targeting Vector Design

    • Strategies for Introducing Subtle Mutations

    • Target Cells for Homologous Recombination

    • Site-Specific Recombination Systems

    • Applications of Site-Specific Recombination

    • Gene Silencing Using Antisense RNA Constructs

    • Gene Silencing Using Inhibitory Protein Strategies

    • Epigenetic Transient Gene Silencing with Oligonucleotides

    • Strategies for Conditional Gene Silencing

  • The Production of Transgenic Mammals from Manipulated Cells

    • Overview

    • Pronuclear Microinjection

    • Variations on Pronuclear Microinjection

    • Embryonic Stem Cells

    • Primoridal Germ Cells

  • Uses for Transgenic Animals

    • Overview

    • Studies Addressing Gene Function

    • Knockout (Null) Mice

    • Studies Addressing Gene Regulation

    • Disease Models

    • Nonmammalian Species as Heterologous Systems

    • Expression Modeling

    • Transgenic Livestock

    • Modulation of Production Traits

    • Animal Bioreactors

  • Nuclear Transfer Technology

    • Overview

    • Advantages of Cell-Based Gene Transfer Procedures

    • Ruminant Embryonic Cells

    • Nuclear Transfer

    • Source of Donor Nuclei

    • Reprogramming

    • Xenotransplantation

    • Nuclear Transfer and Somatic Cell Therapy

    • Animal Cloning

  • References

Keywords:

  • genetic engineering;
  • animal cell;
  • gene transfer;
  • recombinant DNA;
  • vectors