Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
Open Access
- 28 October 2019
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Communications
- Vol. 10 (1), 1-12
- https://doi.org/10.1038/s41467-019-12922-y
Abstract
The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we achieve protein and SpCas9 or AsCas12a ribonucleoprotein (RNP) delivery to cultured human well-differentiated airway epithelial cells and mouse lungs with engineered amphiphilic peptides. These shuttle peptides, non-covalently combined with GFP protein or CRISPR-associated nuclease (Cas) RNP, allow rapid entry into cultured human ciliated and non-ciliated epithelial cells and mouse airway epithelia. Instillation of shuttle peptides combined with SpCas9 or AsCas12a RNP achieves editing of loxP sites in airway epithelia of ROSA(mT/mG) mice. We observe no evidence of short-term toxicity with a wide-spread distribution restricted to the respiratory tract. This peptide-based technology advances potential therapeutic avenues for protein and Cas RNP delivery to refractory airway epithelial cells.Funding Information
- U.S. Department of Health & Human Services | NIH | National Heart, Lung, and Blood Institute (1UG3 HL-147366-01, P01 HL-51670, P01 HL-091842)
- U.S. Department of Health & Human Services | NIH | National Heart, Lung, and Blood Institute
- U.S. Department of Health & Human Services | NIH | National Heart, Lung, and Blood Institute
- U.S. Department of Health & Human Services | NIH | National Institute of Diabetes and Digestive and Kidney Diseases (P30 DK-54759)
- Cystic Fibrosis Foundation (MCCRAY15XX0)
- Roy J. Carver Charitable Trust
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