Engineering branched ionizable lipid for hepatic delivery of clustered regularly interspaced short palindromic repeat-Cas9 ribonucleoproteins

The delivery of the CRISPR/Cas ribonucleoprotein (RNP) has received attention for clinical applications owing to its high efficiency with few off-target effects. Lipid nanoparticles (LNPs) are potential non-viral vectors for the delivery of RNPs. Herein, we report the engineering of a branched scaff...

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Bibliographic Details
Published iniScience Vol. 27; no. 10; p. 110928
Main Authors Onuma, Haruno, Shimizu, Rina, Suzuki, Yuichi, Sato, Mina, Harashima, Hideyoshi, Sato, Yusuke
Format Journal Article
LanguageEnglish
Published Elsevier Inc 01.10.2024
Elsevier
Subjects
Bioengineering
Biological sciences
Drug delivery system
Nanoparticles
Bioengineering
Nanoparticles
Biological sciences
Drug delivery system
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Summary:The delivery of the CRISPR/Cas ribonucleoprotein (RNP) has received attention for clinical applications owing to its high efficiency with few off-target effects. Lipid nanoparticles (LNPs) are potential non-viral vectors for the delivery of RNPs. Herein, we report the engineering of a branched scaffold structure of ionizable lipids for the hepatic delivery of RNPs. Both the total carbon number and branching position were critical for the functional delivery of RNPs. The optimal ionizable lipid exhibited a more than 98% reduction in transthyretin protein after a single dose with no obvious signs of toxicity. The mechanistic study has revealed that optimal LNPs have a unique “flower-like structure” that depends on both the lipid structure and the payload and that these LNPs accumulate in hepatocytes in an apolipoprotein E-independent manner. These results represent a major step toward the realization of in vivo genome editing therapy via RNP delivery using chemically synthesizable LNP formulations. [Display omitted] •Structure of ionizable lipid suitable for hepatic delivery of Cas9 RNP is identified•The RNP-loaded LNP achieved over 98% reduction of TTR protein level after a single dose•The best ionizable lipid is biodegradable and provides excellent safety for the LNPs•The finding supports the feasibility of in vivo genome editing therapy by RNP delivery Bioengineering; Biological sciences; Drug delivery system; Nanoparticles
ISSN:2589-0042
2589-0042
DOI:10.1016/j.isci.2024.110928