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Insect Genetic Technologies ABSA 2017 IBBR-ITF David O’Brochta, Ph.D. Institute for Bioscience and Biotechnology Research Department of Entomology University of Maryland, College Park [email protected] Why, How Biosafety Challenges

Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

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Page 1: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Insect Genetic Technologies

ABSA 2017

IBBR-ITFDavid O’Brochta, Ph.D.Institute for Bioscience and Biotechnology ResearchDepartment of EntomologyUniversity of Maryland, College [email protected]

Why, How

Biosafety Challenges

Page 2: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Insect Genetic Technologies:

NOW

Page 3: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Why?Research Applications

Functional Genomics Physically Contained

Genetically Contained

Uncontained

Page 4: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

ResearchFunctional Genomics

SporozoiteSalivary Gland

Saglin

CRISPR/Cas9 Mutants

Page 5: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Physically Contained Applications

Genetically Modified Mosquitoes and a Malaria Vaccine

Sanaria, Inc.

Page 6: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Genetically Modified Mosquitoes and a Malaria Vaccine

Create Immune-compromised Mosquitoes

Transposon-based Transgenics

Binary Gene Expression System

Page 7: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Genetically Contained Applications

Oxitec, LLC

RIDL StrategyRelease of Insects with Dominant Lethals

Transposon-based Transgenics

Conditional Promoter

run-away transcription

Page 8: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Uncontained Applications

Gene Drive Systems

Page 9: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Gene Drive Systems

Erradication:

Sex-Ratio DistortionReduce Fitness

Gene Introgression:

Anti-parasite genesAnti-virus genes

Page 10: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

General Strategy for Genetically Modifying Insects

X

Page 11: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Delivery of Genetic Technologiesto Insect Germ Cells

Embryo Microinjection

Page 12: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Aedes aegypti

Page 13: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Delivery by Early-Embryo Microinjection Limits Availability of Technologies

Page 14: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Early-Embryos Inaccessible in Many Species

Page 15: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Technical Challenges Limiting the Use of Insect Genetic Technologies

RearingGeneticsScreeningLine Establishment

Delivery

Page 16: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

X

General Strategy for Genetically Modifying Insects

Page 17: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

For Most Insect ScientistsInsect Genetic Modification

is a Heavy Lift

Page 18: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

The Insect Transformation Facility

Since 2006

Page 19: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

The Insect Transformation Facility

Insect Genetic Modification Services:Transposon-based technologiesSite specific recombination technologiesEndonuclease-based ‘gene editing’

TrainingConsulting

Since 2006

Page 20: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

The Insect Transformation Facility

Since 2006

400+ Projects55 Projects in FY2017

40+ Species

Page 21: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

The Insect Transformation Facility

Since 2006

Common Species:Aedes aegypti

Anopheles stephensAnopheles gambiae

Tribolium castaneumDrosophila virilis

Drosophila simulans

Common Technologies:Transposons

Site-specific RecombinationCRISPR/Cas

Page 22: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

The Insect Transformation Facility

Since 2006

Recent Species:Bemisia tabaci

Phlebotomous papatasiAnopheles arabiensis

Anopheles funestisHermetia illucensIxodes scapularisSitobion avenae

Rhopalosiphum padiGryllus bimaculatus

Challenges:What’s Regulated?

Containment requirements?Shipping requirements?

Page 23: Insect Genetic Technologies Why, How · Embryo Microinjection. Aedes aegypti. Delivery by Early-Embryo Microinjection Limits Availability of Technologies . Early-Embryos Inaccessible

Insect Genetic Technologies

ABSA 2017

IBBR-ITFDavid O’Brochta, Ph.D.Institute for Bioscience and Biotechnology ResearchDepartment of EntomologyUniversity of Maryland, College [email protected]

Why, How

Biosafety Challenges