Delivery of CRISPR/Cas9 mRNA LNPs to Repair a Small Deletion in FVIII Gene in Hemophilia A Mice
Highlights From the ASGCT 25th Annual Meeting

Delivery of CRISPR/Cas9 mRNA LNPs to Repair a Small Deletion in FVIII Gene in Hemophilia A Mice

Presented by: Chun-Yu Chen, PhD, Fellow-PhD | Immunity & Immunotherapies, Seattle Children's Seattle, Washington, United States

Chun-Yu Chen1, Cai Xiaohe1, Carol Miao1,2

1Seattle Children’s Research Institute, Seattle, Washington

2Department of Pediatrics, University of Washington, Seattle, Washington

Key Data Points

mRNA LNPs Target Both Hepatocytes and Liver Endothelial Sinusoidal Cells (LSECs)

Various formulations of luciferase mRNA (Luc) encapsulated in MC3-based lipid nanoparticles (LNPs) were intravenously injected into mice. Luciferase expression was mainly detected in liver, with the strongest expression shown by LNP formulation 5. Liver immunostaining staining demonstrated that Dil-labeled LNP can successfully transfect both hepatocytes and endothelial cells in vivo.

Cas9 and sgRNA LNPs Enable NSG HemA Mice to Regain Endogenous Plasma FVIII Activity

Encapsulated Cas9 mRNA and single-guide RNA (sgRNA) LNPs were delivered into immunodeficient hemophilia A (NSG HA) mice intravenously. Stable expression of endogenous FVIII for up to 4 weeks was confirmed by aPTT assay and edited mutant FVIII was verified by DNA sequencing and online CRISPR analysis tools, respectively.

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Supported by educational grants from Bayer, BioMarin, Freeline Therapeutics Limited, Pfizer Inc., Shire, Spark Therapeutics, and uniQure, Inc.

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