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Fast and Efficient CRISPR/Cas9 Genome Editing In Vivo Enabled by Bioreducible Lipid and Messenger RNA Nanoparticles
Open accessAbstract
A main challenge to broaden the biomedical application of CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat (CRISPR) associated protein 9) genome editing technique is the delivery of Cas9 nuclease and single‐guide RNA (sgRNA) into the specific cell and organ. An effective and very fast CRISPR/Cas9 genome editing in vitro and in vivo enabled by bioreducible lipid/Cas9 messenger RNA (mRNA) nanoparticle is reported. BAMEA‐O16B, a lipid nanoparticle integrated with disulfide bonds, can efficiently deliver Cas9 mRNA and sgRNA into cells while releasing RNA in response to the reductive intracellular environment for genome editing as fast as 24 h post mRNA delivery. It is demonstrated that the simultaneous delivery of Cas9 mRNA and sgRNA using BAMEA‐O16B knocks out green fluorescent protein (GFP) expression of human embryonic kidney cells with efficiency up to 90%. Moreover, the intravenous injection of BAMEA‐O16B/Cas9 mRNA/sgRNA nanoparticle effectively accumulates in hepatocytes, and knocks down proprotein convertase subtilisin/kexin type 9 level in mouse serum down to 20% of nontreatment. The leading lipid nanoparticle, BAMEA‐O16B, represents one of the most efficient CRISPR/Cas9 delivery nanocarriers reported so far, and it can broaden the therapeutic promise of mRNA and CRISPR/Cas9 technique further.
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“Gene-editing components can be delivered by lipid nanoparticle–mRNA formulations to treat genetic diseases, including HTI92,238, hypercholesterolaemia81,96,105,239, lipoprotein metabolism disorders85 and transthyretin amyloidosis239,240, which has been demonstrated in preclinical studies.”
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Rinoldi, C., Zargarian, S. S., Nakielski, P., Li, X., Liguori, A., Petronella, F., Presutti, D., Wang, Q., Costantini, M., Sio, L. D., et al. (2021) · Small Methods · doi:10.1002/smtd.202100402 · Cited by 78
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Lipid Nanoparticles: A Novel Gene Delivery Technique for Clinical Application
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“Pathogenic genes have also been corrected using Cas9 nuclease delivered by LNPs [24,31,32].”
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Harnessing lipid nanoparticles for efficient CRISPR delivery
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Hashiba, A., Toyooka, M., Sato, Y., Maeki, M., Tokeshi, M., & Harashima, H. (2020) · Journal of Controlled Release · doi:10.1016/j.jconrel.2020.08.031 · Cited by 66
Nanotechnology Approaches for Chloroplast Biotechnology Advancements
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“CRISPR/Cas9 genome editing in mammalian cells through mRNA delivery has been demonstrated over the span of months (Liu et al., 2019).”
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Non-viral strategies for delivering genome editing enzymes
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CRISPR–Cas9 delivery strategies for the modulation of immune and non-immune cells
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Nanobiomaterial vectors for improving gene editing and gene therapy
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Lipid- and polymer-based nanoparticle systems for the delivery of CRISPR/Cas9
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Stimuli-responsive nanoformulations for CRISPR-Cas9 genome editing
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“Reproduced with permission [129] (See figure on next page.”
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Opportunity and challenges for nanotechnology application for genome editing in plants
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CRISPR/Cas9 Delivery Systems to Enhance Gene Editing Efficiency
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“Lipid-based systems, for example, use cationic lipids to form stable lipoplexes that enhance the payload protection, cellular uptake, and endosomal escape, ultimately improving the genome editing efficiency [21,22].”
InfluentialMasarwy, R., Stotsky‐Oterin, L., Elisha, A., Hazan‐Halevy, I., & Peer, D. (2024) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2024.115359 · Cited by 59
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“With regards to in vivo utility, carboxylated mannan (Man-COOH) was further designed to coat the BPAE-SS/siMOF + miR21 (BSsm) NCs, which served to enhance the serum stability of NCs via shielding of surface cationic charges and to facilitate the internalization of NCs by the mannose receptor-expressing macrophages [49…”
Background
Metadata
- Authors
- Liu, J., Chang, J., Jiang, Y., Meng, X., Sun, T., Mao, L., Xu, Q., & Wang, M.
- Affiliations
- Chinese Academy of Sciences (CN)
- Jilin University (CN)
- Tufts University (US)
- Corresponding author
- Xu, Qiaobing
- Wang, Ming
- Venue
- Advanced Materials
- Publisher
- Wiley
- Published
- 2019-06-19
- Volume
- 31
- Issue
- 33
- Pages
- e1902575-e1902575
- ISSN
- 0935-9648 (ISSN-L)
- 1521-4095
- PMC
- PMC6732788
- OpenAlex
- W2950978761
- Cited by
- 373
- References
- 33
- Citations by year
- 2026: 33 · 2025: 52 · 2024: 53 · 2023: 55 · 2022: 74 · 2021: 53 · 2020: 45 · 2019: 8
- FWCI
- 14.88
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Advanced Materials (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Basic Research Program of China (2017YFA0208100, 2016YFA0200104)
- National Natural Science Foundation of China (21778056, 21790390, 21790391, 21621062, 21435007)
- National Science Foundation (UG3 TR002636‐01, NIH R21 EB024041)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Keywords
- CRISPR/Cas9 genome editing, lipid nanoparticles, hepatocyte targeting, in vivo gene editing
- MeSH terms
- Proprotein Convertase 9, Gene Editing, CRISPR-Associated Protein 9, RNA, Guide, CRISPR-Cas Systems, Animals, Biological Transport, Humans, Lipids, Oxidation-Reduction, RNA, Messenger, RNA, Guide, Kinetoplastida, Gene Transfer Techniques, Cell Line, Tumor, Green Fluorescent Proteins, Mice, Nanoparticles, Gene Knockdown Techniques
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