Transposagen Genome Engineering Brochure 2014

FindYour Place in Genome Engineering
Transposagen Biopharmaceuticals, Inc. 1-844-GEN-EDIT
info@transposagenbio.com
www.transposagenbio.com
Technologies,Tools & Services Brochure
piggyBac™ • XTN™ TALENs • NextGEN™ CRISPR • Footprint-Free™ Gene Editing
• Custom Vectors • Cell Line Engineering • Custom Animal Models and More!
Connect with us for the latest updates on gene editing
technologies, products, services and promotions!

Transposagen Biopharmaceuticals, Inc. www.transposagenbio.com
About Us
Welcome to Transposagen Biopharmaceuticals, Inc., a worldwide leader in genome engineering technologies and
services with applications in therapeutics, research & drug discovery, bioproduction, clinical genetic testing and
agriculture. Our products and services include Footprint-Free™ Gene Editing, NextGEN™ CRISPR, XTN™ TALENs,
and custom cell lines, stem cells, and animal models. Our unique genome engineering capabilities allow for the
creation of nearly any genetic modification in any genome.
• Anyone can cut DNA (even site-
specifically)
• Our technology is the cleanest
gene editing technology in the
world
• We are the only company in the
world that can edit as little as a
single nucleotide without any
unwanted changes and with the
ability to select for rare events
• We have a 20-year patent
position protecting this
technology
Custom reagents and
technology portfolio
Cell and
animal model
engineering
services
Drug development
partnerships
What Differentiates Us WhatWe Offer
• Drug Discovery & Development
Elucidating gene function & regulation
Target discovery
Disease models
Toxicology & reporter lines
Isogenic control lines & diagnostics
• Bioproduction
Host engineering for improved
production and function
Stable cell lines and pools
• Therapeutics
Gene therapy
Cell therapy – ex vivo cell therapy
Why Genome Engineering
1-844-GEN-EDIT

The piggyBac™DNA Modification System
Transposagen is proud to provide the industry’s most efficient non-viral stable transfection & transgenesis
technology. Choose any of our off-the-shelf transposons and stable transfection kits or custom transposon services
in order to facilitate stable transfection and transgenesis. Simple, one step protocol from transfection to stable cell
lines.
How ItWorks
Step 1. Clone your custom cargo into
piggyBac™ transposon vector.
Step 2. Transfect the piggyBac transposon
containing inserted cargo along with the
piggyBac™ transposase (plasmid DNA or
mRNA) into target cell.
Step 3. piggyBac™ transposase recognizes
PB transposon sequences (ITR’s), cuts
transposon, carrying the cargo out of the
plasmid and integrates it stably in the
genome.
Advantages
 No size limit- up to 250 kb has been published
 80-100% stable integration efficiency
 Effective on all mammalian genome
 Precise Excision
 Non- Toxic and non-mutagenic
Applications
• Stable cell line creation
• Stem cell research: reprogramming, differentiation
and selection.
• Protein production
• Cell and gene therapy
• Drug Screens
• RNAi: inducible and reversible knockdown in cells and
animals models
Using piggyBac™ for Stable Transfection & Transgenesis
PB- GOLD™ Stable Transfection Kit Contents
• hyperactive super piggyBac™
transposase (sPBo) plasmid
• lipofection reagents
• positive control plasmid
Unmatched Stable Expression
THP-1 Reporter Lines
Red = negative control
Green = initial transfection expression
Blue = expression after 10 months
Mossine et al. 2013
Outperforms Viral & Plasmid Methods
1-844-GEN-EDIT

piggyBac™ Excision-only Transposase
piggyBac™ excision only transposase allows you to
remove cargo from the in a Footprint-Free™, seamless or
scarless manner. Use for:
 Footprint-Free Gene Editing
 Phenotype Reversion Studies
Custom Services
Transposagen provides everything from vector to full cell
line or animal model creation.
• Custom Transposon vectors
• Custom Cell Lines
• Custom Transgenic animal models
piggyBac™ Footprint-Free™ Excision
PB-x GOLD™ Excision-only Transfection Kit
Contents
• Excision-only piggyBac™ transposase
(PBx) plasmid
• lipofection reagents
• positive control plasmid
RapidTransgenesis
Pettitt et al 2013
Genetic Screen’s & Phenotype Reversion Studies
Demonstration of the piggyBac™
capability to produce large stable
mutant libraries in forward genetic
gene trapping screens. Genes
involved in compound (Olaparib)
toxicity resistance were discovered
(left). Followed by phenotype
reversion back to sensitivity with
excision only piggyBac™ (right)
Green = wild type sensitive cells
Blue = resistant cells with PB gene trap insertions
Grey = reverted excised clones
Blue= resistant cells with gene trap insertion
Green = wild type sensitive cells
Red = negative control
Off-the-shelf vectors
Transposon backbone vectors for cloning your cargo of
interest. The piggyBac™ ITRs flanking core insulators and a
multiple cloning site (MCS). Our backbones come with a
variety of promoters, markers and other features.
1-844-GEN-EDIT

Site-Specific Nucleases for Gene Editing
Site-specific nucleases enable researchers to accurately cut any DNA sequence, allowing for the creation of
gene knock-outs, knock-ins and single base pair edits. Transposagen offers the widest variety of nucleases
including our XTN™ TALENs and the latest dimeric RNA-guided NextGEN™ CRISPR. Not only can Transposagen
provide you with the latest DNA editing technologies, but we have the fastest delivery time in the industry.
Genome Modification Applications
Avoid Off-Targets With NextGEN™ CRISPR
Some nucleases, especially the original CRISPR/Cas9
technology have the unfortunate consequence of off-
target mutations. The NextGEN™ CRISPR technology is the
first truly dimeric RNA-guided FokI nuclease which
diminishes these concerns. The NextGEN™ CRISPR system
empowers researchers to engineer the genome with the
highest fidelity without compromising the budget.
 Affordable
 Fast
 High efficiency
 Able to multiplex
 Highest fidelity; no detectable off-
targets by deep sequencing
methods
• Gene knock-out and knock-ins
• Single base-pair gene editing
• Mutation correction or creation
• Cell line & animal model engineering
• Validation of GWAS studies and patient
clinical trial selection
• Cellular therapies
NextGEN™ architecture. The NextGEN™ CRISPR utilizes the dimerization requirement of FokI for improved specificity.
By introducing site-specific double-strand breaks,
nucleases promote the NHEJ and HR pathways which
enable gene editing.
Custom Reagents
1-844-GEN-EDIT

Succeed With XTN™ TALENs
Transposagen is an experienced XTN™ TALEN producer. In fact, we use XTN™ TALENs for our own in-house
projects for producing genetically engineered rats and cells lines. Our XTN™ TALENs have to be of the highest
quality and activity in order to achieve results for our customers. We deliver our commercial XTN™ TALENs with
the same standards in order to help you achieve your gene editing goals.
Get the best – Not all TALENs are equal.
Transposagen uses the superior protein
engineering to deliver high performance TALENs.
Enhanced targeting, better promoters, variable
repeats and codon optimization make all the
difference.
Have them fast – We utilize our own XTN™ high-
throughput production system to deliver your
TALENs quickly.
Target your gene more often – XTN™ TALENs have
a wide targeting range in the genome making all
types of research possible.
Acquire results – Higher efficiencies translate to
better results.
Stay out of trouble – XTN™ TALENS have minimal
off-target effects and low toxicity.
Site-Specific Nuclease Product & Service Options
Transposagen has a number of innovative site-specific nuclease service programs designed to fit the needs and budget
of any research or drug discovery project. We offer everything from off-the-shelf gene editing tools to the most
complex animal model creation services.
Advantages of XTN™TALENs
→ Value™ Program – Receive custom NextGEN™ CRISPR and/or XTN™ TALEN plasmids sequenced verified and
transfection ready.
→ Guaranteed Success™ Program – Two custom NextGEN™ CRISPR and/or XTN™ TALEN plasmids sequenced
verified and transfection ready. If the first pair of nucleases do not cut at your target you build more for FREE.
→ Gene Editing Kit – Two custom NextGEN™ CRISPR and/or XTN™ TALEN plasmids and custom donor vector
sequenced verified and transfection ready.
→ mRNA Production Service – High quality mRNA production for any custom NextGEN™ CRISPR and/or XTN™
TALEN.
TakeYour Nucleases ToThe Next Level
Ask your local Gene Editing Specialist about our unique Footprint-Free™ Gene Editing piggyBac™ reagents and services.
Combine NextGEN™ CRISPR With XTN™ TALENs
Unlike other providers in the industry, Transposagen offers both CRISPR and TALEN technologies to researchers. Since
each nuclease has inherent sequence requirements, the option of both provides a wider targeting range than alone.
Combine both nucleases in a single transfection for even higher chances of successful gene targeting!
Custom Reagents
XTN™ TALEN Rat Rosa26 targeting compared
to a competitor TALEN producer.
XTN™ Competitor
Bands indicate
activity by Cel1
assay
Evidence of XTN™ Efficiencies
• Hundreds of successful projects in cell line and animal model
creation.
• Direct comparisons between our XTN™ TALENs and competitor
producers.
1-844-GEN-EDIT

Footprint-Free™ Gene Editing
Transposagen’s Footprint-Free™ gene editing kits & services combine the industry’s most precise, efficient, and
flexible site-specific nucleases, NextGEN™ CRISPRs and XTN™ TALENs, with our exclusive piggyBac™ transposon
system - the only commercially available method capable of seamless excision of resistance or reporter genes.
Footprint-Free™ Gene Editing allows users to increase efficiencies by utilizing selection enrichment for gene edits,
followed by “footprint-free” or “scarless” removal of the selection cassette, resulting in clean genome editing.
How ItWorks
1- Start with the most efficient &
precise targeted nucleases,
NextGEN™ CRISPR and/or XTN™
TALENs to stimulate homologous
recombination at the target site.
2- Use a Footprint-Free™ Gene
Editing donor plasmid from our
Multivector™ portfolio for
targeted integration and selection
enrichment of the desired gene
edit.
3- Following selection Excision
Only piggyBac™ (PBx) removes
markers, leaving behind a scarless
correctly edited genomic DNA.
Genome Modification Applications
• Gene editing down to a single base-pair –
mutation correction or creation in cell lines with
key advantages in difficult to transfect cells such
as stem cells (iPS and ES cells) and primary cells.
• Targeted knock-ins, reporter lines and
humanizations.
• Targeted knockouts and knockout reversions –
introduce a disruption (selection marker) in the
coding region, following phenotypic analysis
remove the disruption Footprint-Free™ for
phenotype reversion studies.
Product & Service Options
Transposagen provides a range of services from the off-
the-shelf Multivector™ portfolio to custom gene editing
kits. Footprint-Free™ cell lines and animal model
services are also available.
Multivector™ Catalog #
PB-MV1Puro-TK
PB-MV1Neo-TK
PB-MV1CMV-GFP-Puro
PB-MV1Neo
PB-MV1Hygro-TK
Technology Applications
1-844-GEN-EDIT

Project Applications
Cell line engineering - Access to cell lines and stem cells from genetic disease
or modified backgrounds is becoming more common and important for in vitro
(cell based) modeling. Genomes vary in population, including SNPs in coding
regions and regulatory elements as well as copy number. When compared to
“control or regular stem cells” these natural variations result in
misinterpretation of in vitro phenotypes, making creation of isogenic lines used
as a reference for genetically modified or disease lines necessary for quality
results.
Advantages in Stem Cells - Genome engineering in common transfection cell
lines such as HEK293 occurs at high efficiencies. However, stem cells are less
amendable to genome modification due to reduced transfection efficiencies
and potentially different DNA repair responses. Therefore, selection for
genome modifications through Footprint-Free™ Gene Editing is a key benefit
for stem cell engineering.
Cellular therapies – treatments for a broad range of disorders with clinical
capabilities (no off-target mutations). piggyBac™ and Footprint-Free™ Gene
Editing have been used in CAR T-cell therapy studies as well as correction of
multiple disease mutations.
 Isogenic cell line
creation
 GWAS Study
Validation
 Target discovery &
validation
 Patient clinical trial
selection
 Cell & GeneTherpay
 Animal model
creation
Selection vs. single-stranded DNA oligonucleotide
(ssODN) mediated gene editing: One alternative to
selection is the use of ssODNs which are typically 60-120
bases with the donor substitution (base-pair edits)
flanked by homologous sequences on each side.
Inefficiencies require a large number of clones to be
screened resulting in project timeline delays and cost
increases. Footprint-Free™ selection enrichment allows
simpler easier screening, for faster and more cost
effective genome editing.
Advantages of Footprint-Free™ Gene Editing
Why selection? Site-specific nucleases increase the efficiency of targeted knock-ins or gene edits significantly, but
in many cases efficiencies may not be high enough for clone isolation. Therefore, the ability to select for desired
mutations and rare events is important for efficient gene editing.
Footprint-Free™ Gene Editing vs. CRE/loxP
recombinase: Recombinase methods remove drug
selection markers from the genome by catalyzing
recombination between target sites (known as loxP
sites). However, this process is not clean and leaves
“footprints” or “scars” in the genome which have
been shown to disturb splicing elements or
alter/silence gene expression. In addition,
recombinases recognize “cryptic” loxP sites and
produce off-target mutations, compromising results.
Footprint-Free™ gene editing does not have these
adverse effects providing precise and efficient
genome editing for quality results.
Access to Footprint-Free™
Gene Editing
1- Multivector™ Portfolio – generate
your own Footprint-Free™ gene editing
vectors
2- Custom Footprint-Free™ Gene
Editing Kits – Let us design and
produce the vectors for you
3- Custom Cell Line Engineering &
Animal Model Creation – Utilize
our expertise and outsource the
whole project
Technology Applications
1-844-GEN-EDIT

Custom Cell Line Engineering
Transposagen utilizes expertise and a diverse portfolio of gene editing tools including XTN™ TALENs, NextGEN™
CRISPRs, piggyBac™ transposons and Footprint-Free™ Gene Editing to deliver high quality custom engineered cell lines
Available Services
 Stable cell lines (transgenic,
overexpression, inducible)
 Knockout
 Knock-in (humanization &
conditional)
 Footprint-Free™ Gene Editing
 Cell line characterization services
Custom Services
An example of our 4-step milestone pricing structure for
high efficiency knockout cell lines
Best Reagents
Your custom engineered cell line begins with the latest gene editing
reagents available. Transposagen uses TALEN and CRISPR
technologies, including the latest dimeric NextGEN™ CRISPR which has
been shown to have minimal off-target effects. Combined with
piggyBac Footprint-Free Gene Editing, Transposagen achieves the
highest efficiency.
Expertise
Transposagen has over 10 years of experience engineering custom cell
lines and animal models. We've successfully delivered projects to the
top 20 pharmaceutical companies and all the major academic
institutes. We have experience with many cell lines including iPS, SSCs,
CHO, HEK293, HeLa, ES, SH-SY5Y, HepG2, A549, MCF7, CACO-2,
RAW264.7, THP-1, Jurkat... to name a few!
Time
Generating a cell line can be time-consuming
and complex. By utilizing the piggyBac™ system,
Transposagen can delivery engineered cell lines
faster than anyone in the industry for even the
most difficult lines.
Accurate Pricing & Flexible Projects
No surprises. Prior to the start of any project,
Transposagen can accurately price out the cost
of your engineered cell line so you know what to
expect. All cell line projects are divided into
milestone payments which allows researchers to
start the service without paying for all of it
upfront. You only pay for the work we complete!
KeepYour Investment
The reagents we use to engineer your cell line
are yours to keep. Use them again in a different
cell line or take them to our animal model
services to maximize your research efforts.A demonstration of knockout
cell line creation at
Transposagen. Inactivating
the fucosylation system in
CHO cells for producing
afucosylated human
therapeutics. Antibodies
expressed from these host
cells were afucosylated and
demonstrated the expected
increased ADCC activity.
In collaboration with Aragen
Bioscience.
Targeting Fut8, the alpha1,6-fucosyltransferase gene
with XTN™TALENs in CHO Cells
1-844-GEN-EDIT

Custom Animal Models
Transposagen has the industry’s most versatile collection of genome engineering technologies as well as
expertise in rodent pronuclear injection (PNI) and stem cell engineering. These capabilities result in the most
flexible and robust custom model generation services in the industry. The more targeted and complicated your
therapeutics become, the more you need the right partner to advance your research. Working intimately with
our scientific team will help you bring innovative solutions and products to market faster.
Custom Services
Available Rat & Mouse Services
 Knockout
 Transgenic Overexpression
 Conditional
 Knock-in
 Transgenic knockdown
 Humanized
 Disease Models
1-844-GEN-EDIT
Genome Engineering Vector Design & Validation
+
PNI, ES or SSCs
High-speed, high-efficiency genome engineering
Rodent Model Creation Strategies
Method Species Targeted Knock-in
Limitation
Estimated Timeline
Pronuclear Injection (PNI) Mouse & Rat ~2kb 4 months
Embryonic Stem Cells (ES) Mouse None 8 months
Spermatogonial Stem
Cells (SSC)
Rat None 8 months
• Custom Engineered Rodents in as little as 4 months
• Spermatogonial Stem Cell (SSC) technology is the only commercial method for producing
large targeted knock-ins in rats

FatRat™ Mc4r Knockout Obesity-Diabetes Model
Research Models
Featured Knockout Rat Models
Drug Metabolism
• Bcrp
Immune System
• Rag2 - SCID
Neuroscience –CNS
• SERT
• OPRL1
• Ghsr
• Nrg1
• Trpc4
• Pde4d
• Myo9a
Metabolic Disease/Obesity
• Mc4r – FatRat™
• Leptin
Cardiovascular
• Sod3
Cancer/Toxicology
• P53
1-844-GEN-EDIT
Transposagen’s off the shelf TKO® and TGEM® rat models cover a range of disease and research applications. We
have partnered with world experts in over 40 collaborations worldwide in order to validate and characterize our
genetically engineered rat models. We have approximately 300 off-the-shelf models which is the world’s largest
commercially available collection. Our characterized genetically engineered models are broken down into the
research categories below.
• Most common monogenic obesity gene
• Obesity studies, including early and late onset obesity
• Diabetes research
• Metabolism studies
• Renal studies
• Altered Leptin Signaling Pathway
Rag2 –SCID Knockout Rat
• Cell sorting data demonstrates no mature T
and B cells
• Western blot /Immunochemistry indicates
no IgG
• Initial results indicate that the animals are
athymic
New SCID Rat for Human Xenograft
Tumor Models
Access to Research Models
Unlimited breeding licenses - breeding at the
customer’s in-house facility or a contract breeder of
choice. The license comes with at least 2-3
heterozygous breeding pairs.
Trial licenses- allow the customer to purchase a
limited breeding license to conduct pilot studies at
a lower purchase price.
Individual animals – may be available, please
inquire

Transposagen Biopharmaceuticals, Inc. 1-844-GEN-EDIT
www.transposagenbio.com
Connect with us for the latest updates on gene editing
technologies, products, services and promotions!
Exon Exon Exon
One-Step Stable
Transfection & Transgenesis
Targeted Knockouts
Footprint-Free™
Gene Editing
NextGEN™ CRISPR
StableTransfection Kits &
piggyBac™ Transposon
XTN™TALEN
Knock-ins &Humanization
Gene Editing
Base-Pair Changes
Cell Line & Rodent Model Engineering Services
Your Next Discovery Awaits
Targeted Selection
Markers
With So Many Opportunities
What AreYou Waiting For?
Next-Generation Genome Engineering

Transposagen Genome Engineering Brochure 2014

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