Cover

Contents

Contributors

Acknowledgements

Preface

PART 1: GERMLINE TRANSFORMATION TECHNOLOGY

1 Transposons for Insect Transformation

1.1 Transposable Elements

1.2 DNA Transposons

1.3 Transposons with Activity in Insects

1.4 Summary

References

2 Transposon-Based Technologies for Insects

2.1 Transposon-Based Technologies

2.2 Mutagenesis

2.3 Germ-Line Transformation

2.4 Modular Expression Systems

2.5 Cell/Genetic Ablation

2.6 Transgene Mis-expression

2.7 Transgenic Gene Silencing

2.8 Site-Specifi c Recombination

2.9 Genetic Sensors

2.10 Conclusions

References

3 Sex-, Tissue- and Stage-Specifi c Transgene Expression

3.1 Introduction

3.2 Gene Regulation in Insects

3.3 Th e Basic Genetic Construct

3.4 Controlling for Position Eff ects

3.5 General Considerations for Promoter Choice

3.6 Sex-Specifi c Transgene Expression

3.7 Tissue-Specifi c Expression

3.8 Stage-Specifi c Expression

3.9 Future Prospects

3.10 Applications of Sex-, Tissue- and Stage-Specifi c Transgene Expression

References

4 Docking Systems for Site-Directed Transgene Integration

4.1 Background and Introduction

4.2 Site-Specifi c Transgenesis – Generation of Phase 1 Docking Strains

4.3 Site-Specifi c Transgenesis – Generation of Phase 2 Integrations

4.4 Recombinase-Mediated Cassette Exchange

4.5 Future Developments in the Use of Docking Systems

4.6 Docking Systems Combined with Transposon Stabilization Strategies

4.7 Integration of Large, Complex Transgene Constructs

4.8 Construction of Complex Transgenes by Sequential Use of Integrases

References

5 Inducible and Repressible Systems for Transgene Expression

5.1 Introduction

5.2 Naturally Occurring Systems of Conditional Expression

5.3 Synthetic Systems

5.4 Conclusions

References

6 Sex Ratio Manipulation for Insect Population Control

6.1 Introduction

6.2 Overview of Applications and General Principles

6.3 Meiotic Drive

6.4 Sex-Specifi c Lethality

6.5 Manipulation of Sex Determination Mechanisms

6.6 Conclusions

References

7 Conditional Dominant Lethals – RIDL

7.1 Re-engineering the Sterile Insect Technique

7.2 Sterile Insects and Genetic Control

7.3 Engineered Traits

7.4 Integrated Pest Management

7.5 Resistance Management

7.6 Molecular Designs

7.7 Choosing an Eff ector

7.8 Choice of Switch

7.9 Strain Performance

7.10 Penetrance

7.11 Resistance

7.12 Field Experience and Future Prospects

Notes

Acknowledgements

References

PART 2: APPLICATIONS OF TRANSGENIC INSECTS

8 Tephritid Fruit Fly Transgenesis and Applications

8.1 Introduction

8.2 Transformation with the Minos Vector System

8.3 Transformation with the piggyBac Vector System

8.4 Transformation with the Hermes Vector System

8.5 Marker Systems for Transformant Organismal and Tissue Detection in Tephritid Flies

8.6 Post-integration Stabilization of Transposon Vectors in Tephritid Flies

8.7 Site-Specifi c Genomic Targeting in Tephritids

8.8 Transgenic Strains for Improved Population Control of Tephritids

Acknowledgement

References

9 Silkworm Transgenesis and Applications

9.1 Introduction

9.2 Generation of Transgenic Silkworms

9.3 Application of Transgenic Silkworms to Gene Function Analyses

9.4 Production of Recombinant Proteins for Pharmaceutical Use

9.5 Construction of Modifi ed Silk and its Possible Use as a Biomaterial

9.6 Gene Targeting

9.7 Future Prospects

References

10 Transgenic Approaches for Sterile Insect Control of Dipteran Livestock Pests and Lepidopteran Crop Pests

10.1 A Brief History of Using the Sterile Insect Technique for Controlling Populations of Agricultural Pests

10.2 Enhancing the Sterile Insect Technique Th rough Transgenic Technologies: an Overview

10.3 Enhancing the Sterile Insect Technique Th rough Transgenic Technologies: New World Screwworm and the Australian Sheep Blowfl y

10.4 Enhancing the Sterile Insect Technique Th rough Transgenic Technologies: Lepidoptera

10.5 Future Directions

Acknowledgements

References

11 Antipathogen Eff ector Molecules: Current and Future Strategies

11.1 Introduction

11.2 Eff ector Molecules

11.3 Manipulating Mosquito Physiology: Insulin Signalling as a Case Study for Modifying Immunity, Lifespan and Reproduction

11.4 Conclusions

References

12 Sexual Sterilization of Mosquitoes

12.1 Introduction

12.2 Genetic Sterility Versus Irradiation

12.3 Spermless Males Induce Life-Long Sterility in Females

12.4 Genetic Sterility Th rough the Expression of Testis-Specifi c Eff ector Genes

12.5 Targeting the Function of the Male Accessory Glands

12.6 Male Sterility Genes: What Is Known in Drosophila

12.7 Biotechnology Toolbox to Generate Sterility

12.8 Disrupting Fertility by Classical Transgenesis

12.9 Target Gene Disruption by Homologous Recombination-Based Gene Knock-Out or Replacement

12.10 Gene Knock-Out Using Synthetic Endonucleases

12.11 Culturing Sexually Sterile Mosquito Lines 201 Acknowledgements

Acknowledgements

References

PART 3: ALTERNATIVE TRANSGENIC APPROACHES TO MODIFYING INSECT PHENOTYPES

13 Paratransgenesis in Mosquitoes and Other Insects: Microbial Ecology and Bacterial Genetic Considerations

13.1 Introduction

13.2 Requirements for Successful Paratransgenesis

13.3 Paratransgenesis of Mosquitoes Against Malaria With Genetically Modifi ed Bacteria

13.4 Paratransgenesis With Naturally Occurring Bacterial Strains

13.5 Conclusions

Acknowledgements

References

14 Asaia Paratransgenesis in Mosquitoes

14.1 Asaia in Mosquitoes

14.2 Asaia and Paratransgenesis in Mosquito-Borne Disease Control

14.3 Asaia is Capable of Cross-Colonizing Insects of Diff erent Genera and Orders

14.4 Asaia Within Mosquitoes: What Are its Benefi cial Roles?

14.5 Future Perspectives

References

15 Paratransgenic Control of Chagas Disease

15.1 Introduction

15.2 Chagas Disease

15.3 Novel Approaches to Eradication of Chagas Disease

15.4 From Bench Top to Field Trials

15.5 Conclusions

References

16 Tsetse Paratransgenesis: a Novel Strategy for Reducing the Spread of African Trypanosomiasis

16.1 Tsetse as Vectors of Parasitic African Trypanosomes

16.2 Tsetse Symbiosis – Transmission Routes and Functions

16.3 Tsetse Paratransgenesis

16.4 Taxonomic Characterization of the Tsetse Microbiome

16.5 Mechanisms to Drive Parasite-Resistant Tsetse Phenotypes into Natural Populations

16.6 Conclusions

References

PART 4: CONSIDERATIONS FOR THE RELEASE OF TRANSGENIC INSECTS

17 RIDL: Modelling Release of Insects Carrying a Dominant Lethal

17.1 Sterile Insect Methods

17.2 A Genetic Twist

17.3 It’s the Ecology, Stupid!

17.4 Th e Aim of the Game

17.5 All Together Now

17.6 Follow the Money

17.7 Wish List

References

18 Assessing Risk of Transgenic Insects

18.1 Introduction

18.2 Risk Assessment

18.3 Risk Assessment of Living Insects

18.4 Risk Assessment of Genetically Modifi ed Organisms

18.5 Special Aspects of Risk for Transgenic Insects

18.6 Documentation of Risk Assessment

18.7 Social and Political Aspects of Risk

18.8 Conclusions

Notes

Acknowledgements

References

19 Economics of Transgenic Insects for Field Release

19.1 Introduction

19.2 Inundative Concept

19.3 Inoculative Concept

19.4 Funding Investment and Capturing Benefi ts

19.5 Capturing Public Health Benefi ts

19.6 Conclusions

References

20 Risk Analysis and the Regulation of Transgenic Insects

20.1 Introduction

20.2 Genetic Engineering

20.3 Common Features of Regulatory Systems

20.4 Regulatory Gaps and Overlaps

20.5 Conclusions

Notes

References

21 Public Acceptability of New Insect Vector Control Technologies

21.1 Introduction

21.2 Th e On-Going Challenge of Vector Control

21.3 Th e Need for Alternative Public Health Strategies to Control Vector-Borne Diseases

21.4 Th e New Technologies

21.5 Challenges For Th e Public Acceptability of New Vector Technologies

21.6 Mechanisms to Address Challenges for Public Engagement

21.7 Community Engagement

21.8 Informed Consent

21.9 Conclusions

References

22 Th e Cartagena Protocol on the Transboundary Movement of Living Modifi ed Organisms: Th e Regulation of Trade in Transgenic Organisms under International and European Environmental Law

22.1 Introduction

22.2 Overview of the UN Convention on Biological Diversity

22.3 Cartagena Protocol on Biosafety (2000/2003)

22.4 Conclusions

Notes

References

Index

A

B

C

D

E

F

G

H

I

J

L

M

N

O

P

Q

R

S

T

U

V

W

X

Y

Z