Systemic biodistribution and hepatocyte-specific gene editing with CRISPR/Cas9 using hyaluronic acid-based nanoparticles
| Autor: | Mansoor M. Amiji, C.M. Francis, Liliana Wroblewska, Pamela Pegman |
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| Rok vydání: | 2021 |
| Předmět: |
Biodistribution
Biomedical Engineering Pharmaceutical Science Medicine (miscellaneous) Mannose Bioengineering Green fluorescent protein chemistry.chemical_compound Mice Genome editing Hyaluronic acid medicine Animals General Materials Science Tissue Distribution Hyaluronic Acid Gene Editing technology industry and agriculture Transfection Molecular biology medicine.anatomical_structure chemistry Hepatocyte Systemic administration Hepatocytes Molecular Medicine Nanoparticles CRISPR-Cas Systems |
| Zdroj: | Nanomedicine : nanotechnology, biology, and medicine. 40 |
| ISSN: | 1549-9642 |
| Popis: | The goal of this study was to evaluate hepatocyte-specific gene editing, via systemic administration of hyaluronic acid (HA)-based nanoparticles in naive CD-1 mice. Using HA-poly(ethylene imine) (HA-PEI) and HA-PEI-mannose nanoparticles with differential mannose density (1X and 2X), we have evaluated systemic biodistribution and hepatocyte-specific delivery using IVIS imaging and flow cytometry. Additionally, we have investigated hepatocyte-specific delivery and transfection of CRISPR/Cas9 gene editing plasmid and eGFP gene payload to integrate at the Rosa26 locus. IVIS imaging showed uptake of HA-PEI nanoparticles primarily by the liver, and with addition of mannose at different concentrations, the nanoparticles showed increased uptake in both the liver and spleen. HA-PEI-mannose nanoparticles showed 55–65% uptake by hepatocytes, along with uptake by resident macrophage regardless of the mannose concentration. One of two gRNA targets showed 15% genome editing and obtained similar results for all three nanoparticle formulations. Cells positive for our gene payload were greatest with HA-PEI-mannose-1X nanoparticles where 16.2% of cells were GFP positive. The results were encouraging as proof of concept for the development of a non-viral biodegradable and biocompatible polymeric delivery system for gene editing specifically targeting hepatocytes upon systemic administration. |
| Databáze: | OpenAIRE |
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