CRISPR Therapeutics : ASGCT 2024 Oral Presentation

May 8, 2024 | ASGCT, ORAL SESSION, ABSTRACT #87

Development of an In Vivo Non-Viral Delivery Platform for Ocular Editing and Application as a Potential Treatment for Glaucoma

Mary-Lee Dequéant, Ph.D. | Senior Director, CRISPR Therapeutics

Forward-Looking Statements

The presentation and other related materials may contain a number of "forward-looking statements" within the meaning of the Private Securities Litigation Reform Act of 1995, as amended, including statements regarding CRISPR Therapeutics' expectations about any or all of the following: (i) its plans and expectations for its preclinical studies, clinical trials and pipeline products and programs; (ii) the safety, efficacy and clinical progress of its various clinical programs; (iii) the status of preclinical studies and clinical trials (including, without limitation, the expected timing of data releases, announcement of additional programs and activities at clinical trial sites, and discussions with regulatory authorities) and expectations regarding the data that is being presented; (iv) the data that will be generated by ongoing and planned preclinical studies and clinical trials and the ability to use that data for the design and initiation of additional preclinical studies and clinical trials; (v) regulatory submissions and authorizations, including timelines for and expectations regarding additional regulatory agency decisions; (vi) manufacturing activities and capabilities; (vii) the activities under its collaborations and the expected benefits thereof; (viii) its intellectual property coverage and positions of its, its licensors and third parties as well as the status and potential outcome of proceedings involving any such intellectual property; (ix) the sufficiency of its cash resources; and (x) the therapeutic value, development, and commercial potential of CRISPR/Cas9 gene editing technologies and therapies, including as compared to other therapies. Without limiting the foregoing, the words "believes," "anticipates," "plans," "expects" and similar expressions are intended to identify forward-looking statements. You are cautioned that forward-looking statements are inherently uncertain. Although CRISPR Therapeutics believes that such statements are based on reasonable assumptions within the bounds of its knowledge of its business and operations, forward-looking statements are neither promises nor guarantees and they are necessarily subject to a high degree of uncertainty and risk. Actual performance and results may differ materially from those projected or suggested in the forward-looking statements due to various risks and uncertainties. These risks and uncertainties include, among others, that: the efficacy and safety results from ongoing clinical trials will not continue or be repeated in ongoing or planned clinical trials or may not support regulatory submissions; regulatory authorities may not approve exa-cel on a timely basis or at all; adequate pricing or reimbursement may not be secured to support continued development or commercialization of exa-cel following regulatory approval; the potential that clinical trial results may not be favorable; one or more of its product candidate programs will not proceed as planned for technical, scientific or commercial reasons; future competitive or other market factors may adversely affect the commercial potential for its product candidates; initiation and completion of preclinical studies for its product candidates is uncertain and results from such studies may not be predictive of future results of future studies or clinical trials; regulatory approvals to conduct trials or to market products are uncertain; it may not realize the potential benefits of its collaborations; uncertainties regarding the intellectual property protection for its technology and intellectual property belonging to third parties, and the outcome of proceedings (such as an interference, an opposition or a similar proceeding) involving all or any portion of such intellectual property; and those risks and uncertainties described under the heading "Risk Factors" in its most recent annual report on Form 10-K, quarterly report on Form 10-Q, and in any other subsequent filings made by it with the U.S. Securities and Exchange Commission, which are available on the SEC's website at www.sec.gov. Existing and prospective investors are cautioned not to place undue reliance on these forward-looking statements, which speak only as of the date they are made. CRISPR Therapeutics disclaims any obligation or undertaking to update or revise any forward-looking statements contained in this presentation, other than to the extent required by law.

Caution should be exercised when interpreting results from separate trials involving separate product candidates. There are differences in the clinical trial design, patient populations, and the product candidates themselves, and the results from the clinical trials of some product candidates or products (e.g., autologous product candidates or products) may have no interpretative value on our existing or future results.

CRISPR THERAPEUTICS® standard character mark and design logo, CTX110®, CTX112 , CTX130 , CTX131 , CTX310 , CTX320 , CTX330 and CTX211 are trademarks and registered trademarks of CRISPR Therapeutics AG. All other trademarks and registered trademarks are the property of their respective owners. Solely for convenience, trademarks, service marks and trade names referred to in this presentation may appear without the ® or symbols and any such omission is not intended to indicate waiver of any such rights.

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Disclosures

Mary-Lee Dequéant, Ph.D.

Senior Director, CRISPR Therapeutics

Disclosure Information

I have the following relevant financial relationships to disclose: Employee of: CRISPR Therapeutics

Stockholder in: CRISPR Therapeutics

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Non-Viral Delivery - A Promising Approach

for Ocular Gene Editing

● Potential to use CRISPR/Cas9 technology to address

genetic diseases of the eye once considered untreatable	Lipid nanoparticle (LNP) formulated with
	Cas9 mRNA and guide RNA (gRNA)

• Advantages of localized delivery to the eye:

• Accessible for localized delivery

○ Small volume

• • Enclosed structure / limited systemic exposure
  • Immune-privilegedorgan
• Non-viraldelivery of Cas9 mRNA offers several advantages:
• • Transient expression of Cas9 enzyme, no potential for vector integration
  • Ability to deliver larger cargoes
  • Simplified manufacturing
  • Repeat dosing feasible

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MYOC Mutations are the Most Common

Genetic Cause of Glaucoma

• Glaucoma is the second leading cause of blindness worldwide1
• Myocilin (MYOC) mutations are the most common genetic cause of glaucoma.2 MYOC-associated glaucoma affects ~150k people in the United States alone1,3
• Myocilin mutations are autosomal dominant4 and associated with earlier onset and more rapid progression of the disease5
• Mutations cause impaired outflow of aqueous humor through the trabecular meshwork (TM), leading to elevated intraocular pressure (IOP)6

Abnormal aqueous humor outflow

through the TM Increased intraocular pressure (IOP)

Damage to optic nerve and ganglion cells

1 Allingham R.R. et al. 2009, 2 Fingert J.H. et al., 2011, 3 Zhang N. et al., 2021, 4 Sheffield V.C. et al. 1993, 5 Fingert J.H. et al., 1999,

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6 Alward W.L.M. et al. 1998.

Myocilin Gene is a Promising Target for the Treatment of Glaucoma

• Myocilin is a glycoprotein highly expressed in the trabecular meshwork (TM)1
• Gain-of-functionMYOC mutations cause misfolded protein aggregates in the endoplasmic reticulum (ER), inducing cellular stress that can lead to TM cell death/dysfunction2
• Knocking out MYOC expression in the TM using CRISPR/Cas9 was shown to reverse the ER stress, restore TM function, and reduce IOP3,4
• WT MYOC expression is not required for normal ocular function5 and homozygous MYOC loss of function in humans is not associated with any disease6

Adapted from Kasetti et al., 2021

1 Pang C.P. et al. 2002, 2 Kasetti R.B. et al. 2016, 3 Jain A. et al., 2017, 4 Patil S.V. et al., 2024, 5 Kim B.S. 2001,	CRISPR THERAPEUTICS	6
6 Lam D.S. 2000.

In Vivo Screen Identified LNPs that Deliver Efficiently to Mouse TM

• ~200 LNPs formulated with EGFP1 mRNA were tested in vitro in primary TM cells

● ~40 LNP formulations were selected for in vivo testing	EGFP Detected in TM by IHC3
24h After LNP Dosing

EGFP Expression in Mouse TM 24h After IVT2 Injection		Trabecular
3		Meshwork
Score
2
GFP 1	Ciliary body	Iris
0	100μm
LNP Formulations

1 EGFP - Enhanced Green Fluorescent Protein; 2 IVT - Intravitreal; 3 IHC - Immunohistochemistry

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LNPs Selected for Targeted Delivery to Non-

Human Primate TM

Intracameral Injection	EGFP Detected in TM by IHC
in Cynomolgus Monkey	24h After Dosing with LNP
	Iris
	TM

Ciliary

Body

Cornea

Injection in the anterior segment of LNP formulated with EGFP mRNA

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LNP Formulation Also Successfully

Delivered to Human TM in Ex Vivo Model

Human Eye Anterior Segment	EGFP Detected in Human TM by IHC
Organ Culture (ASOC)1	24h After LNP Infusion

TM

200μm

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1 LNP formulated with EGFP mRNA was infused through syringe/pump. Flow: 2.5 μl/min (method adapted from Boice and Snider, JOVE, 2021)

Prioritized MYOC gRNA Shows Efficient

Editing in Primary Human TM cells

>90% MYOC Gene Editing In Vitro	MYOC Protein Reduction Up to 85%
with Prioritized gRNA1	with Prioritized gRNA

Editing (%)

100

80

60

40

20

0

No LNP

Low

Medium

High

LNP Dose

Protein Expression	(norm.)
MYOC

1.0

0.8

0.6

0.4

0.2

0.0

No LNP

Low

Medium

High

LNP Dose

Cross-reactive human/NHP MYOC gRNA selected based on in vitro screen

Comprehensive off- target assessment completed with no off-target editing detected for prioritized gRNAs

1 gRNA - guide RNAs	CRISPR THERAPEUTICS	10