1. CRISPR-Cas Representing Abundant Potential for Agriculture
Nandini Krishnamurthy,
Senior Research Manager, Genome Edited Trait Discovery –
Trait Discovery & Optimization, DuPont Pioneer

2. DuPont Ag’s key technology platforms
BIOTECH
BREEDING
CRISPR
SEED APPLIED
TECHNOLOGY
CROP PROTECTION
DIGITAL SOLUTIONS
AGRONOMIC SOLUTIONS
“We put a great emphasis in managing our grain on a much more micro level – so looking not on a field by field basis but more on an acre by acre and even in some cases on a sub-acre level.”
- DuPont Customer
Only DuPont can collaborate with growers acre by acre to answer these demands. Everything we do is about solving one problem – a complex problem that has grown and changed with the needs of our growing world –
CROP PRODUCTIVITY

3. Why CRISPR-Cas?
Efficient and targeted plant breeding technology Enables customized solutions for farmer’s challenges
Don’t believe that this slide is needed

4. Conventional Breeding Practices Involve Many Generations of Crossing
Drought tolerance example:
100% A
50%:50%
25%:75%
12.5%:87.5%
6.75%:93.25%
3.38%:96.62%
Tolerant to drought
Lower quality plant
Sensitive to drought
Higher quality plant
GOAL
100% high quality plant +
drought tolerance
100% B
Traditional plant breeding involves many crosses of parental lines to achieve the product targets within our elite germplasm. This is an example of one characteristics. Complexity increases with each additional characteristic.

5. CRISPR-Cas Enables Efficient Introduction of Desired Characteristics
All corn plants should only have one ear, change in all slides
CRISPR-Cas Enables Efficient Introduction of Desired Characteristics
Drought tolerance example:
FROM: Incorporating desired characteristics in multiple cycles of common breeding practices TO: Incorporating desired characteristics in as little as1 to 2 cycles via CRISPR- Cas advanced breeding
Drought sensitive
Desired characteristic
Drought tolerant
100% A
100% B
GOAL
100% high quality plant +
drought tolerance
Lower quality plant
Tolerant to drought
Higher quality plant
Sensitive to drought
Suggest that this slide follows the conventional breeding slide

6. How Can We Use CRISPR-Cas?
DELETE
The plant does not have desired output.
The plant does have desired output.
The plant has tolerance to drought.
The plant has high tolerance to drought.
EDIT
The plant is susceptible to disease.
The plant is resistant to disease.
SEARCH/REPLACE
CRISPR-Cas is a flexible tool with many applications
It works similar to how a copy editor proofs and edits an article. CRISPR-Cas looks at the plant’s native attributes, much like a copy editor reads the first draft of an article. Then, CRISPR-Cas adds, removes or repositions attributes native in the plant to achieve a characteristic like drought or disease tolerance. This is similar to how a copy editor improves a narrative through word choice. CRISPR-Cas works with what naturally exists within the crop’s available characteristics to making the resulting plants thrive under difficult growing conditions.
The delete application was applied to create our next generation of waxy corn hybirds – which will get into in more detail a bit later.

7. DNA Breaks and Repair Happen in Nature
INTERNAL STRESS
DNA BREAK
DNA REPAIR
EXTERNAL STRESS

8. CRISPR-Cas Enables Targeted DNA Breaks
Cas9 nuclease (DNA cutting enzyme)
Guide RNA is designed to direct Cas9 enzyme to the DNA sequence of interest
Cas9 enzyme binds to the targeted DNA and makes double strand break
Target Sequence
DNA double strand break is repaired through a plant’s own cellular process
Guide RNA
Guide RNA matches the target DNA sequence
CRISPR-Cas Applications 4

9. Pioneer Will Deploy Targeted Breeding Broadly
NEAR-TERM PRODUCTS TO MARKET
BROAD AGRICULTURAL APPLICATIONS OF CRISPR-CAS
WAXY CORN HYBRIDS
Foundational for future product development
First commercial agricultural product
To market by end of current decade
DISEASE RESISTANCE
YIELD & YIELD STABILITY
DROUGHT TOLERANCE
OUTPUT TRAITS
MATURITY
CORN •
SOY
CANOLA
RICE
WHEAT
SUNFLOWER
NORTHERN CORN LEAF BLIGHT
Devastating global disease with potential to cause $1.6B* annual losses in North America alone
Leveraging germplasm base
Utilizing native genes, genomic selection, and genome editing
Providing sustainable grower solutions
NORTHERN LEAF BLIGHT READINESS: First half of next decade
Products, benefits and concepts described herein will not be offered for sale or distribution until completion of field testing and applicable regulatory reviews. * Source: Internal analysis and USDA.

10. Waxy Cornstarch is Part of Everyday Consumer Products
Essential functions in food industry to improve uniformity, stability, and texture in various food products Binding qualities for the paper- making process Additional applications in the, corrugating, textile and adhesive industries

11. More information available
CRISPRCas.pioneer.com