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Review
. 2021 Jul 12;22(14):7456.
doi: 10.3390/ijms22147456.

Exosome/Liposome-like Nanoparticles: New Carriers for CRISPR Genome Editing in Plants

Mousa A Alghuthaymi  1 , Aftab Ahmad  2   3 , Zulqurnain Khan  4 , Sultan Habibullah Khan  3 , Farah K Ahmed  5 , Sajid Faiz  6 , Eugenie Nepovimova  7 , Kamil Kuča  7 , Kamel A Abd-Elsalam  8
Affiliations
Review

Exosome/Liposome-like Nanoparticles: New Carriers for CRISPR Genome Editing in Plants

Mousa A Alghuthaymi et al. Int J Mol Sci. .

Abstract

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.

Keywords: CRISPR; exosomes and liposomes; genome editing; nanoparticles.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Delivery methods of CRISPR/Cas cargos into plants. (A) Biolistic transformation is based on the delivery of DNA into plant cells by high-velocity gold or tungsten particles with the help of a gene gun; (B) protoplast transformation involves the direct delivery of DNA into individual plant cells using polyethylene glycol or electroporation; (C) Agrobacterium-mediated transformations utilize the capability of the bacterial pathogen Agrobacterium tumefaciens to transfer foreign genes into a wide variety of host plants; (D) Agrobacterium rhizogene transformations transport single-stranded DNA (ssDNA; T-strands) and virulence proteins into plant cells through a type IV secretion system; (E) agroinfiltration transformations allow the suspension culture of agrobacterial cells to be infiltrated into the organs of an intact plant, providing a rapid and efficient way to transiently express foreign genes in planta.
Figure 2
Figure 2
CRISPR/Cas9 cargos for delivery into cells. (A) Cas9 protein. (B) Trans-CRISPR RNA, sgRNA, and the Cas9–sgRNA complex can be delivered into cells using different methods. Some of the important methods are shown in the figure. (C) Cas9 protein delivery; (D) Cas9-expressing cell lines; (E) delivery of ribonucleoprotein (RNP); (F) delivery in the viral vectors/plasmids; (G) Oligos; (H) Delivery of the crRNA/tracrRNA complex as mRNA.
Figure 3
Figure 3
CRISPR/Cas reagents: (A) traditional Cas9 reagent; (B) CRISPR/Cas base editor; (C) prime editor.
See this image and copyright information in PMC

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