A fluorescent splice-switching mouse model enables high-throughput, sensitive quantification of antisense oligonucleotide delivery and activity.

Cell reports methods 2024 Jan 22

filter terms:

While antisense oligonucleotides (ASOs) are used in the clinic, therapeutic development is hindered by the inability to assay ASO delivery and activity in vivo. Accordingly, we developed a dual-fluorescence, knockin mouse model that constitutively expresses mKate2 and an engineered EGFP that is alternatively spliced in the presence of ASO to induce expression. We first examined free ASO activity in the brain following intracerebroventricular injection revealing EGFP splice-switching is both ASO concentration and time dependent in major central nervous system cell types. We then assayed the impact of lipid nanoparticle delivery on ASO activity after intravenous administration. Robust EGFP fluorescence was observed in the liver and EGFP+ cells were successfully isolated using fluorescence-activated cell sorting. Together, these results show the utility of this animal model in quantifying both cell-type- and organ-specific ASO delivery, which can be used to advance ASO therapeutics for many disease indications. Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.

Citation

Amy E Byrnes, Filip Roudnicky, Alvin Gogineni, Allison L Soung, Monica Xiong, Margaret Hayne, Tiffany Heaster-Ford, Whitney Shatz-Binder, Sara L Dominguez, Jose Imperio, Sarah Gierke, Jasmine Roberts, Jinglong Guo, Soumitra Ghosh, Charles Yu, Merone Roose-Girma, Justin Elstrott, Amy Easton, Casper C Hoogenraad. A fluorescent splice-switching mouse model enables high-throughput, sensitive quantification of antisense oligonucleotide delivery and activity. Cell reports methods. 2024 Jan 22;4(1):100673