Enhanced Cytosolic Delivery and Release of CRISPR/Cas9 by Black Phosphorus Nanosheets for Genome Editing

Zhou, Wenhua; Cui, Haodong; Ying, Liming; Yu, Xue‐Feng

doi:10.1002/ange.201806941

Cited by 59 publications

(40 citation statements)

References 40 publications

(21 reference statements)

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“…Several studies that have successfully employed nanoparticle delivery of CRISPR-Cas9 plasmids have done so using immortalized cell lines [31][32][33]. In this way, our study is unique as we utilize primary cells.…”

Section: Discussionmentioning

confidence: 99%

Fabrication and characterization of PLGA nanoparticles encapsulating large CRISPR–Cas9 plasmid

Ringel‐Scaia

McDaniel

et al. 2020

J Nanobiotechnol

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Background: The clustered regularly interspaced short palindromic repeats (CRISPR) and Cas9 protein system is a revolutionary tool for gene therapy. Despite promising reports of the utility of CRISPR-Cas9 for in vivo gene editing, a principal problem in implementing this new process is delivery of high molecular weight DNA into cells. Results: Using poly(lactic-co-glycolic acid) (PLGA), a nanoparticle carrier was designed to deliver a model CRISPR-Cas9 plasmid into primary bone marrow derived macrophages. The engineered PLGA-based carriers were approximately 160 nm and fluorescently labeled by encapsulation of the fluorophore 6,13-bis(triisopropylsilylethynyl) pentacene (TIPS pentacene). An amine-end capped PLGA encapsulated 1.6 wt% DNA, with an encapsulation efficiency of 80%. Release studies revealed that most of the DNA was released within the first 24 h and corresponded to ~ 2-3 plasmid copies released per nanoparticle. In vitro experiments conducted with murine bone marrow derived macrophages demonstrated that after 24 h of treatment with the PLGA-encapsulated CRISPR plasmids, the majority of cells were positive for TIPS pentacene and the protein Cas9 was detectable within the cells. Conclusions: In this work, plasmids for the CRISPR-Cas9 system were encapsulated in nanoparticles comprised of PLGA and were shown to induce expression of bacterial Cas9 in murine bone marrow derived macrophages in vitro. These results suggest that this nanoparticle-based plasmid delivery method can be effective for future in vivo applications of the CRISPR-Cas9 system.

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Section: Discussionmentioning

confidence: 99%

Fabrication and characterization of PLGA nanoparticles encapsulating large CRISPR–Cas9 plasmid

Ringel‐Scaia

McDaniel

et al. 2020

J Nanobiotechnol

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“…To address this issue, researchers recently have shown tremendous enthusiasm for the march toward the exploration of nonviral vectors ( 4 ). To date, various vehicles, including gold nanoparticles ( 12 ), black phosphorus ( 13 ), metal-organic frameworks ( 14 ), graphene oxide ( 15 ), polymeric nanoparticles ( 16 , 17 ), and other nanomaterials ( 18 , 19 ), have been achieved availably to deliver CRISPR-Cas9 in vitro or in vivo. However, on-demand release of CRISPR-Cas9 for the precise remote temporal and spatial control over genome editing remains elusive.…”

Section: Introductionmentioning

confidence: 99%

Near-infrared upconversion–activated CRISPR-Cas9 system: A remote-controlled gene editing platform

et al. 2019

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As an RNA-guided nuclease, CRISPR-Cas9 offers facile and promising solutions to mediate genome modification with respect to versatility and high precision. However, spatiotemporal manipulation of CRISPR-Cas9 delivery remains a daunting challenge for robust effectuation of gene editing both in vitro and in vivo. Here, we designed a near-infrared (NIR) light–responsive nanocarrier of CRISPR-Cas9 for cancer therapeutics based on upconversion nanoparticles (UCNPs). The UCNPs served as “nanotransducers” that can convert NIR light (980 nm) into local ultraviolet light for the cleavage of photosensitive molecules, thereby resulting in on-demand release of CRISPR-Cas9. In addition, by preparing a single guide RNA targeting a tumor gene (polo-like kinase-1), our strategies have successfully inhibited the proliferation of tumor cell via NIR light–activated gene editing both in vitro and in vivo. Overall, this exogenously controlled method presents enormous potential for targeted gene editing in deep tissues and treatment of a myriad of diseases.

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“… 110 Apart from solid lipid nanoparticles, many other novel nanoparticles have also been investigated for CRISPR-Cas delivery: gold nanoclusters, 111 gold nanowires, 112 nanoscale zeolitic imidazole frameworks, 113 and black phosphorus nanosheets. 114 These novel nanoparticles are just a bourgeoning area of research. Although many nanoparticles have shown promising results in cell culture, their in vivo efficacy and side effects are not very clear, and they often require complex design and it may be difficult to produce high amounts for clinical use.…”

Section: Delivering Methodsmentioning

confidence: 99%

Applications and challenges of CRISPR-Cas gene-editing to disease treatment in clinics

Liu

Jiang

et al. 2021

Precision Clinical Medicine

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Clustered regularly interspaced short palindromic repeats-CRISPR associated systems (Cas) are efficient tools for targeting specific genes for laboratory research, agricultural engineering, biotechnology, and human disease treatment. Cas9, by far the most extensively used gene-editing nuclease, has shown great promise for the treatment of hereditary diseases, viral infection, cancers and so on. Recent reports have revealed that some other types of CRISPR-Cas systems may also have surprising potential to join the fray as gene-editing tools for various applications. Despite the fast progress in basic researches and clinical tests, some underlying problems present continuous, significant challenges, such as editing efficiency, relative difficulty in delivery, off-target effects, immunogenicity, etc. This article summarizes the applications of CRISPR-Cas from bench to bedside and highlights the current obstacles that may limit the usage of CRISPR-Cas systems as gene-editing tool-kits in precision medicine and offer some viewpoints that may help to tackle these challenges and facilitate technical development. CRISPR-Cas systems, as a powerful gene-editing approach, will offer great hopes in clinical treatments for many individuals with currently incurable diseases.

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Enhanced Cytosolic Delivery and Release of CRISPR/Cas9 by Black Phosphorus Nanosheets for Genome Editing

Cited by 59 publications

References 40 publications

Fabrication and characterization of PLGA nanoparticles encapsulating large CRISPR–Cas9 plasmid

Fabrication and characterization of PLGA nanoparticles encapsulating large CRISPR–Cas9 plasmid

Near-infrared upconversion–activated CRISPR-Cas9 system: A remote-controlled gene editing platform

Applications and challenges of CRISPR-Cas gene-editing to disease treatment in clinics

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