Transposagen Q3 2012 Overview

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Better Models. Better Results™

Genetic Modification for Better Disease Models™

© Transposagen Biopharmaceutics, Inc.

About Us

Technologies: • piggyBac™ •XTN™ TAL Target-Specific Nucleases •Rodent Stem Cells

Transposagen Biopharmaceuticals, Inc., is dedicated to providing better disease models through genetic modification. Transposagen specializes in custom and off the shelf XTN™ TAL Nucleases, animal models, cell lines, stem cells and cutting-edge research tools and technologies to improve drug discovery and development research. These unique technologies and services for genetic modification give customers and technology users access to any genetic modification in any gene in any genome for both research and biologic production purposes.

Applications: •Rodent Model Creation •Cell Line Engineering & Stem Cells •Genetically Engineered Plants •Biologics Production •Therapeutics

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piggyBac Site Specific Genetic Engineering

• Not limited to TTAA sites

• Xanthamonas TAL nuclease (XTN: a.k.a TALENs)

• TAL DNA binding modules and FokI DNA-cleavage domain

• Site specific knockout, knock-in of any gene, any genome

• Automated synthesis of XTNs

• Remove transposon after gene editing

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XTN™ TAL Nuclease KO Efficiency

• Rag1 XTN target site amplified, cloned and sequenced

• 16% disruption at the target site: better than Zinc-finger Nuclease and Meganuclease

• Disruption sequences are shown below with XTN target site highlighted in grey

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A. Site specific binding and induced dsDNA breaks

B. Transfect SSCs

C. Nested PCR confirmation of Homologous Recombination events

Site Specific Knock-in

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The piggyBac™ Transposon System

Key Features

•Efficient transgenesis using transfection •Titratable number of integrations •Large cargo limit – 10 to 100kb •Rapid identification of integration site •Reversible transposition – no footprint •All-in-one inducible vector •Safe non-viral system

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piggyBac vs. Viral Vectors

• Cargo capacity greater than 120 kb • Delivery of entire BACs reported

Characteristic Viral Vectors piggyBac

Safety

Induced Immune Response

Oncogene Activation

Production Costs

Cargo Capacity

X

X

X

X

X

Source: Current Gene Therapy (2004), 4, 357-372

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

Cargo Capacity (kb)

piggyBac

Lentivirus

Retrovirus

Adenoassociated Virus

120 kb

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piggyBac™ Cumate Inducible Switch Titratable and can turn back off easily

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Reversible Transposition Pop-out the transposon

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Targeted Integration and Excision

• Gene Therapy • Cell Lines

Animal Models Therapeutics Cell Lines Bradley et al Nature (2011) In Press

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Therapeutic Applications

•Gene and Cell Therapy

•Immunotherapy

PB kills ovarian cancer cells of tumor xenografts in mice

PB/TK cells stained red Apoptotic cells stain green Merged cells are yellow

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Animal Models

• Knockout

• Transgenic Overexpression

• Conditional

• Knock-in

• Transgenic knockdown

• Humanized

Rat and mouse models available Any species possible

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Creation of Animal Models using SSCs

Figures Courtesy of Dr. Kent Hamra

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Genetic Modification Technology Comparison Primary Technology

Virus ZFN & HR Plasmid piggyBac & XTN

High Efficiency Yes No No Yes

Non-viral safety No Yes Yes Yes

Integration control

No Yes No Yes

Unlimited DNA Cargo size

No No No Yes

Reversible footprint free

No No No Yes

Site specific gene editing

No Yes No Yes

Cell lines and Animal Models

Yes Yes No Yes

Licensing restrictions

No Yes No No

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Custom Services Overview • Gene editing with XTNs (talens), HR vector, piggyBac Transposon

Site specific knockout, knock-in, edit and correction of any gene in any genome

• Stable cell lines Overexpression, knockdown, reporter lines, iPS reprogramming, iPS

and ES differentiation and selection, therapeutic applications

• Animal models Rat and mouse model generation available, additional species

possible

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Contact Information

Jack Crawford, MS Director, Business Development jcrawford@transposagenbio.com