Faculty Opinions recommendation of Efficient delivery of genome-editing proteins using bioreducible lipid nanoparticles.

A central challenge to the development of protein-based therapeutics is the inefficiency of delivery of protein cargo across the mammalian cell membrane, including escape from endosomes. Here we report that combining bioreducible lipid nanoparticles with negatively supercharged Cre recombinase or anionic Cas9:single-guide (sg)RNA complexes drives the electrostatic assembly of nanoparticles that mediate potent protein delivery and genome editing. These bioreducible lipids efficiently deliver protein cargo into cells, facilitate the escape of protein from endosomes in response to the reductive intracellular environment, and direct protein to its intracellular target sites. The delivery of supercharged Cre protein and Cas9:sgRNA complexed with bioreducible lipids into cultured human cells enables gene recombination and genome editing with efficiencies greater than 70%. In addition, we demonstrate that these lipids are effective for functional protein delivery into mouse brain for gene recombination in vivo. Therefore, the integration of this bioreducible lipid platform with protein engineering has the potential to advance the therapeutic relevance of protein-based genome editing.

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

International Journal of Molecular Sciences ◽

10.3390/ijms22147456 ◽

2021 ◽

Vol 22 (14) ◽

pp. 7456

Author(s):

Mousa A. Alghuthaymi ◽

Aftab Ahmad ◽

Zulqurnain Khan ◽

Sultan Habibullah Khan ◽

Farah K. Ahmed ◽

...

Keyword(s):

Genome Editing ◽

Viral Vectors ◽

Polymeric Materials ◽

Inorganic Nanoparticles ◽

Plant Genome ◽

Delivery Methods ◽

Detailed Consideration ◽

Crispr System ◽

Efficient Delivery ◽

Low Cytotoxicity

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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Genome Editing in Agriculture: Technical and Practical Considerations

International Journal of Molecular Sciences ◽

10.3390/ijms20122888 ◽

2019 ◽

Vol 20 (12) ◽

pp. 2888 ◽

Cited By ~ 8

Author(s):

Julia Jansing ◽

Andreas Schiermeyer ◽

Stefan Schillberg ◽

Rainer Fischer ◽

Luisa Bortesi

Keyword(s):

Genome Editing ◽

Mechanism Of Action ◽

High Efficiency ◽

Public Acceptance ◽

Crop Species ◽

Base Editing ◽

The Public ◽

Intact Plants ◽

Efficient Delivery ◽

Selection Of

The advent of precise genome-editing tools has revolutionized the way we create new plant varieties. Three groups of tools are now available, classified according to their mechanism of action: Programmable sequence-specific nucleases, base-editing enzymes, and oligonucleotides. The corresponding techniques not only lead to different outcomes, but also have implications for the public acceptance and regulatory approval of genome-edited plants. Despite the high efficiency and precision of the tools, there are still major bottlenecks in the generation of new and improved varieties, including the efficient delivery of the genome-editing reagents, the selection of desired events, and the regeneration of intact plants. In this review, we evaluate current delivery and regeneration methods, discuss their suitability for important crop species, and consider the practical aspects of applying the different genome-editing techniques in agriculture.

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Efficient Delivery of Antisense Oligonucleotides Using Bioreducible Lipid Nanoparticles In Vitro and In Vivo

Molecular Therapy — Nucleic Acids ◽

10.1016/j.omtn.2020.01.018 ◽

2020 ◽

Vol 19 ◽

pp. 1357-1367 ◽

Cited By ~ 5

Author(s):

Liu Yang ◽

Feihe Ma ◽

Fang Liu ◽

Jinjin Chen ◽

Xuewei Zhao ◽

...

Keyword(s):

Antisense Oligonucleotides ◽

Lipid Nanoparticles ◽

Efficient Delivery

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Self-Assembled DNA Nanoclews for the Efficient Delivery of CRISPR-Cas9 for Genome Editing

Angewandte Chemie International Edition ◽

10.1002/anie.201506030 ◽

2015 ◽

Vol 54 (41) ◽

pp. 12029-12033 ◽

Cited By ~ 283

Author(s):

Wujin Sun ◽

Wenyan Ji ◽

Jordan M. Hall ◽

Quanyin Hu ◽

Chao Wang ◽

...

Keyword(s):

Genome Editing ◽

Efficient Delivery ◽

Self Assembled

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Selective RNA interference and gene silencing using reactive oxygen species-responsive lipid nanoparticles

Chemical Communications ◽

10.1039/c9cc04517a ◽

2019 ◽

Vol 55 (56) ◽

pp. 8170-8173 ◽

Cited By ~ 4

Author(s):

Chunjing Liang ◽

Jin Chang ◽

Ying Jiang ◽

Ji Liu ◽

Lanqun Mao ◽

...

Keyword(s):

Gene Expression ◽

Reactive Oxygen Species ◽

Reactive Oxygen ◽

Lipid Nanoparticles ◽

Oxygen Species ◽

Efficient Delivery ◽

Intracellular Ros ◽

Cell Gene Expression ◽

High Level ◽

Cell Gene

The integration of reactive oxygen species (ROS)-responsive thioketal group into lipids nanoparticles enables the efficient delivery of siRNA into cells, and selectively cancer cell gene expression silencing in response to the high level of intracellular ROS in cancer cells.

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Design of Peptidomimetic Functionalized Cholesterol Based Lipid Nanoparticles for Efficient Delivery of Therapeutic Nucleic Acids

Molecules ◽

10.3390/molecules24183413 ◽

2019 ◽

Vol 24 (18) ◽

pp. 3413 ◽

Cited By ~ 1

Author(s):

Ehexige Ehexige ◽

Tsogzolmaa Ganbold ◽

Xiang Yu ◽

Shuqin Han ◽

Huricha Baigude

Keyword(s):

Morphological Changes ◽

Sirna Delivery ◽

Green Fluorescence Protein ◽

Average Diameter ◽

Lipid Nanoparticles ◽

High Yield ◽

Cell Nuclei ◽

Efficient Delivery ◽

Fluorescence Protein ◽

Interfering Rna

Lipid nanoparticles (LNP) are the most potent carriers for the delivery of nucleic acid-based therapeutics. The first FDA approved a short interfering RNA (siRNA) drug that uses a cationic LNP system for the delivery of siRNA against human transthyretin (hTTR). However, preparation of such LNP involves tedious multi-step synthesis with relatively low yields. In the present study, we synthesized cationic peptidomimetic functionalized cholesterol (denote Chorn) in straightforward chemical approaches with high yield. When formulated with helper lipids, Chorn LNPs complexed with siRNA to form nanoparticles with an average diameter of 150 nm to 200 nm. Chorn LNP mediated transfection of a green fluorescence protein (GFP) expressing plasmid resulted in 60% GFP positive cells. Moreover, Chorn LNP delivered siRNA against polo-like kinase 1 (Plk1), a disease related gene in cancer cells and efficiently suppressed the expression of the gene, resulting in significant morphological changes in the cell nuclei. Our data suggested that cholesterol based cationic LNP, prepared through a robust chemical strategy, may provide a promising siRNA delivery system.

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