Exosome/Liposome-like Nanoparticles: New Carriers for CRISPR Genome Editing in Plants
Open accessAbstract
Rapid developments in the field of plant genome editing using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) systems necessitate more detailed consideration of the delivery of the CRISPR system into plants. Successful and safe editing of plant genomes is partly based on efficient delivery of the CRISPR system. Along with the use of plasmids and viral vectors as cargo material for genome editing, non-viral vectors have also been considered for delivery purposes. These non-viral vectors can be made of a variety of materials, including inorganic nanoparticles, carbon nanotubes, liposomes, and protein- and peptide-based nanoparticles, as well as nanoscale polymeric materials. They have a decreased immune response, an advantage over viral vectors, and offer additional flexibility in their design, allowing them to be functionalized and targeted to specific sites in a biological system with low cytotoxicity. This review is dedicated to describing the delivery methods of CRISPR system into plants with emphasis on the use of non-viral vectors.
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Nanotechnology-based approaches including inorganic nanoparticles, carbon nanotubes, liposomes, and protein- and peptide-based nanoparticles offer significant advantages over traditional delivery techniques.
“Innovative nanotechnology-based approaches, including inorganic nanoparticles, carbon nanotubes, liposomes, and protein- and peptide-based nanoparticles, offer significant advantages over traditional delivery techniques.”
From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Alghuthaymi et al., 2021
The Introduction (page unavailable) identifies magnetic nanoparticles and carbon nanotubes as promising for plant transformation, and §4.2 (page unavailable) describes advantages of liposomes for gene delivery. The cited passages do not establish advantages for protein- or peptide-based nanoparticles; they discuss proteins and peptides as potential cargo instead. Nor do they establish that every listed approach is superior to traditional delivery techniques. (LLM, conf=0.86)
Experimental — accuracy not yet measuredNanotechnology-based vectors demonstrate reduced immunogenic responses and lower cytotoxicity.
“These vectors demonstrate reduced immunogenic responses and lower cytotoxicity, making them particularly attractive for genetic modification strategies (Alghuthaymi et al., 2021).”
From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Alghuthaymi et al., 2021
The use of nanoparticles, such as mesoporous silica nanoparticles, liposomes, and layered double hydroxides, as vectors for the delivery of the CRISPR/Cas9 system is a significant development in gene editing.
“The use of nanoparticles, such as mesoporous silica nanoparticles, liposomes, and layered double hydroxides, as vectors for the delivery of the CRISPR/Cas9 system is a significant development in gene editing (Chen and Mueller, 2018;Alghuthaymi et al., 2021).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Alghuthaymi et al., 2021
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BackgroundDelivery of CRISPR/Cas9 for therapeutic genome editing
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RNA-Targeting CRISPR-Cas systems and their applications
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“Gaseous NO, when encapsulated in an echogenic liposome, remains active up to 37 ◦C in the serum, which is suitable for prolonged uses [64].”
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BackgroundCRISPR-Cas12a/Cpf1-assisted precise, efficient and multiplexed genome-editing in Yarrowia lipolytica
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Non-Viral Delivery To Enable Genome Editing
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Efficient gene editing in adult mouse livers via adenoviral delivery of CRISPR/Cas9
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Liposomal delivery of CRISPR/Cas9
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“It could be simply non-specific ionic interaction-mediated endocytosis, or it could be influenced by other factors [142].”
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MethodSolid lipid nanoparticles for applications in gene therapy: a review of the state of the art
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BackgroundExchange of Small Regulatory RNAs between Plants and Their Pests
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BackgroundGenome Editing in Mouse Spermatogonial Stem Cell Lines Using TALEN and Double-Nicking CRISPR/Cas9
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MethodImproved Delivery of CRISPR/Cas9 System Using Magnetic Nanoparticles into Porcine Fibroblast
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Delivery of CRISPR/Cas9 by Novel Strategies for Gene Therapy
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BackgroundCRISPR-induced double-strand breaks trigger recombination between homologous chromosome arms
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“Moreover, Cas9 mRNA and protein formats are superior to those of Cas9 plasmids from the perspective of the reduction of off-target effects, which may induce higher efficiency for double-stranded break (DSB) formation with the help of non-viral carriers.”
BackgroundNanovehicles for Plant Modifications towards Pest- and Disease-Resistance Traits
Kumar, S., Nehra, M., Dilbaghi, N., Marrazza, G., Tuteja, S. K., & Kim, K. (2019) · Trends in Plant Science · doi:10.1016/j.tplants.2019.10.007 · Cited by 65 · cited 1× · see citing works
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Novel cationic solid-lipid nanoparticles as non-viral vectors for gene delivery
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Generation of GGTA1 Mutant Pigs by Direct Pronuclear Microinjection of CRISPR/Cas9 Plasmid Vectors
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“Gene knockout PD-11 Human serum albumin (HSA) nanoparticles [69]”
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BackgroundNanovesicle-Mediated Delivery Systems for CRISPR/Cas Genome Editing
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“For such nanomaterials, techniques and gene delivery approaches continue to be developed, and the key challenge is to balance transfection efficiency, targeting specificity, particle size, biodegradability, and cytotoxicity, as well as their short- and long-term fates in the environment [149].”
BackgroundEfficient lipid-mediated transfection of DNA into primary rat hepatocytes
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“This makes these systems similar to real-life systems [97].”
BackgroundNanobiotechnology in health sciences: Current applications and future perspectives
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“The plant cuticle has amphiphilic properties and small size ranging between 80 and 300 nm [44,50].”
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Highly efficient gene inactivation by adenoviral CRISPR/Cas9 in human primary cells
Voets, O., Tielen, F. J., Elstak, E., Benschop, J., Grimbergen, M., Stallen, J., Janssen, R. A. J., van Marle, A., & Essrich, C. (2017) · PLoS ONE · doi:10.1371/journal.pone.0182974 · Cited by 46 · see citing works
Strategic Nanoparticle-Mediated Gene Transfer in Plants and Animals - a Novel Approach
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BackgroundSelf-citationOptimized nanoparticle-mediated delivery of CRISPR-Cas9 system for B cell intervention
Li, M., Fan, Y.-N., Chen, Z., Luo, Y., Wang, Y., Lian, Z., Xu, C., & Wang, J. (2018) · Nano Research · doi:10.1007/s12274-018-2150-5 · Cited by 43 · cited 1× · see citing works
“Gene knockout BAFFR Polyethyleneimine–cyclodextrin [67]”
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“Liposomes can offer several advantages as vectors for gene delivery into plant cells, including enhanced delivery of encapsulated DNA by membrane fusion, protection of nucleic acids from nuclease activity, targeting to specific cells, and delivery into a variety of cell types besides protoplasts via entry through plas…”
BackgroundLiposome‐mediated gene delivery into plant cells
Gad, A. E., Rosenberg, N., & Altman, A. (1990) · Physiologia Plantarum · doi:10.1111/j.1399-3054.1990.tb05883.x · Cited by 41 · cited 1× · see citing works
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BackgroundCarbon nanotubes: Plant gene delivery and genome editing
Chandrasekaran, R., Periakaruppan, R., & Abd-Elsalam, K. A. (2020) · Elsevier eBooks · doi:10.1016/b978-0-12-819786-8.00014-1 · Cited by 38 · see citing works
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Synthetic Vehicles for Encapsulation and Delivery of CRISPR/Cas9 Gene Editing Machinery
Carboni, V., Maaliki, C., Alyami, M., Alsaiari, S. K., & Khashab, N. M. (2019) · Advanced Therapeutics · doi:10.1002/adtp.201800085 · Cited by 33 · cited 1× · see citing works
“The synthesis of an aptamer–liposome–CRISPR/Cas9 chimaera approach, which led to improved and accurate delivery of CRISPR/Cas9–gRNA in vitro with reduced immunogenicity relative to other cationic liposomal formulations, was facilitated by the modification of liposomal surfaces with aptamers [107].”
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“Nanomaterial-based cell membrane vectors can deliver drugs while keeping their configuration and folding in their native states and maintaining stability [8].”
BackgroundInside-out: from endosomes to extracellular vesicles in fungal RNA transport
Kwon, S., Tisserant, C., Tulinski, M., Weiberg, A., & Feldbrügge, M. (2020) · Fungal Biology Reviews · doi:10.1016/j.fbr.2020.01.001 · Cited by 30 · see citing works
siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes
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“[79] used the transfection reagent lipofectamine in nematodes to improve siRNA uptake from the environment.”
MethodOdahara, M., Watanabe, K., Kawasaki, R., Tsuchiya, K., Tateishi, A., Motoda, Y., Kigawa, T., Kodama, Y., & Numata, K. (2021) · ACS Applied Nano Materials · doi:10.1021/acsanm.1c00695 · Cited by 29 · cited 1× · see citing works
“Biological nanocarriers, such as cell-penetrating peptide-displayed polyion complex vesicles (CPP-PICsomes), have also been used for DNA-free genome editing using Cas9 ribonucleoprotein [151].”
MethodNanoparticles-Based Delivery Systems in Plant Genetic Transformation
Rai, M., Bansod, S., Bawaskar, M., et al. (2015) · Springer eBooks · doi:10.1007/978-3-319-14024-7_10 · Cited by 27 · cited 2× · see citing works
“Gene vectors, constructed to transfer a gene of interest into a host cell, play an important role in genetic transformations [3].”
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“In particular, the use of the lentivirus as a means of delivering DNA to transfection-resistant cells is an effective method for producing stable Cas9 and sgRNA cell lines [18].”
MethodAn Overview of Genetic Transformation of Soybean
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CRISPR/Cas9 Genome Editing Using Gold‐Nanoparticle‐Mediated Laserporation
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“However, it can be used after some modifications [145,146].”
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“Additionally, regulatory oversight for genetically modified foods (GMOs) motivates the development of plant engineering technologies for which gene expression is transient and foreign DNA are not integrated into the host plant genome [1].”
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“Gene knockout Ptch1, Trp53, Pten PEI [72] Gene knockout AAVS1 Gold nanoparticles [73] Gene knockout mCCR7 GNOME [74] Gene knockout CD38 Nano-blade chip [75]”
BackgroundTranscriptomics Evaluation of Hexavalent Chromium Toxicity in Human Dermal Fibroblasts
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“In liposome-based gene therapy there is no potential toxicity for humans and plants [53,54].”
BackgroundPolymer and lipid-based nanoparticles to deliver RNAi and CRISPR systems
Chandrasekaran, R., Seetharaman, P., Danaraj, J., Periakaruppan, R., & Abd-Elsalam, K. A. (2021) · Elsevier eBooks · doi:10.1016/b978-0-12-821910-2.00016-3 · Cited by 9 · cited 1× · see citing works
“The notable benefit of cationic lipid nanoparticles over polymeric NPs is that they can be generated without the use of organic solvents using a high-pressure homogenization (HPH) process [98].”
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Magnetic Nanoparticles: A Unique Gene Delivery System in Plant Science
Mohamed, M. A., & Abd-Elsalam, K. A. (2019) · Nanotechnology in the life sciences · doi:10.1007/978-3-030-16439-3_6 · Cited by 7 · see citing works
Self-citationAdvancements in plant disease control strategies
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Emerging Role of Nanocarriers in Delivery of Nitric Oxide for Sustainable Agriculture
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- Authors
- Alghuthaymi, M. A., Ahmad, A., Khan, Z., Khan, S. H., Ahmed, F. K., Faiz, S., Nepovimova, E., Kuča, K., & Abd-Elsalam, K. A.
- Affiliations
- Shaqra University (SA)
- Center for Advanced Studies in Agriculture and Food Security (PK)
- Muhammad Nawaz Shareef University of Agriculture (PK)
- Cairo University (EG)
- University of Haripur (PK)
- Department of Chemistry, Faculty of Science, University of Hradec Kralove, 50003 Hradec Kralove, Czech Republic
- University of Hradec Králové (CZ)
- Agricultural Research Center (EG)
- Corresponding author
- Kuča, Kamil
- Abd-Elsalam, Kamel A.
- ORCID
- Venue
- International Journal of Molecular Sciences
- Publisher
- MDPI AG
- Published
- 2021-07-12
- Volume
- 22
- Issue
- 14
- Pages
- 7456
- ISSN
- 1422-0067 (ISSN-L)
- PMC
- PMC8304373
- OpenAlex
- W3182877332
- Semantic Scholar
- fa5bda741044430b874d68b05a1df59071579ea7
- Cited by
- 83
- References
- 156
- Citations by year
- 2026: 13 · 2025: 17 · 2024: 11 · 2023: 18 · 2022: 19 · 2021: 5
- FWCI
- 1.39
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- International Journal of Molecular Sciences (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Department of Chemistry, Faculty of Science, University of Hradec Kralove, 50003 Hradec Kralove, Czech Republic (UHK project VT2019-2021)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Transgenic Plants and Applications
- Keywords
- CRISPR/Cas genome editing, plant genome editing, nanoparticle delivery, exosomes, liposomes, non-viral vectors, CRISPR delivery, plant transformation, nanocarriers
- MeSH terms
- Gene Editing, Liposomes, Plants, Genetic Therapy, Genome, Plant, Nanoparticles, Exosomes, CRISPR-Cas Systems
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