1. Gene Editing Tools and Methods
Andrew May DPhil
Chief Scientific Officer
Caribou Biosciences, Inc.
9/18/2018

2. Caribou background
Founded by Jennifer Doudna, Rachel Haurwitz, Martin Jinek and James Berger (UC Berkeley) to develop commercial applications of CRISPR-Cas systems
Co-founded Intellia Therapeutics in 2014 to develop human gene and cell therapies using Cas9
Research partnerships with DuPont, Novartis, Intellia
Located in Berkeley, CA
9/18/2018

3. Caribou CRISPR technology platform is being applied in multiple areas
Gene and cell therapies
Antimicrobials
Animal health
Precision breeding
Novel trait generation
Crop genetics
Cell engineering
Animal models
Phenotypic screening
Genomics
Microbial fermentation
Therapeutic bioproduction
Caribou is pioneering the translation of CRISPR-Cas genome engineering for the development of new bio-based products
9/18/2018

4. We can now translate genomic information into valuable solutions
>5,000 Mendelian disorders understood at the genetic level
>40 crop species with reference genome sequences
>3,400 complete microbial genome sequences
Correct the genetic mutation in somatic cells with gene therapy
Utilize precision breeding to develop crops with improved performance
Selectively kill harmful microbes and preserve ‘friendly’ microbes
9/18/2018

5. Site-directed nucleases for gene editing
Meganucleases
Cellectis, Precision Biosciences, bluebird
Zinc Finger Proteins
Sangamo BioSciences
TALE Nucleases
Cellectis
CRISPR-Cas9
Caribou, Intellia, Editas, CRISPR Tx
Other systems
9/18/2018

6. Therapeutic application areas
Anti-microbial therapies
Infectious disease
Editing microbial genomes
Drug discovery and development
Improved cellular and animal models
Screens for new targets and mechanisms for intervention with small molecules and antibodies
New genomic tools
Antibody, vaccine, viral production
9/18/2018

7. Important considerations for gene editing
Specificity
Where can the nuclease cut?
Where does it cut?
DNA repair outcomes
How does the cell repair the break?
How do these repair outcomes modify target activity?
Functional assays
How do DNA repair outcomes impact function of the cell type of interest?
Disease-relevant models and measurements for assessing activity and safety
9/18/2018

8. SITE-Seq provides comprehensive measurement of Cas9/guide RNA specificity
Comprehensive identification of all possible cut sites in the genome
Measurement of identified sites at high sensitivity allows us to determine activity at each site in cell populations of interest
9/18/2018

9. Analysis of Cas9-dependent double-strand break repair outcomes
9/18/2018

10. Summary
Gene editing technologies hold huge promise for therapeutics and many other fields
Genomic tools potentially enable higher precision activity measurement and detection of effects than for other therapeutic classes
Current regulatory framework has been used successfully to monitor gene editing and gene therapies
Smaller patient populations may require alternative approaches
9/18/2018

11. Gene editing technologies can provide beneficial impact in many different areas
Gene and cell therapies
Antimicrobials
Animal health
Precision breeding
Novel trait generation
Crop genetics
Cell engineering
Animal models
Phenotypic screening
Genomics
Microbial fermentation
Therapeutic bioproduction
Many regulatory considerations are common across these diverse application areas
9/18/2018