Suppressing mosaicism by Au nanowire injector-driven direct delivery of plasmids into mouse embryos

Park, Kkotchorong; Kim, Keun Cheon; Lee, Hyoban; Sung, Yoori; Kang, Mijeong; Lee, Yun Mi; Ahn, Ji Yeon; Lim, Jeong Mook; Kang, Taejoon; Kim, Bongsoo; Lee, Eun Ju

doi:10.1016/j.biomaterials.2017.05.044

Cited by 11 publications

(6 citation statements)

References 75 publications

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“…The plasmids are released through dissociation triggered by an electric pulse. Because the technique is thought to be less violent, embryo survival was reported to be significantly higher than traditional microinjection 677 .…”

Section: Intracellular Delivery By Direct Penetrationmentioning

confidence: 99%

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

2018

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Intracellular delivery is a key step in biological research and has enabled decades of biomedical discoveries. It is also becoming increasingly important in industrial and medical applications ranging from biomanufacture to cell-based therapies. Here, we review techniques for membrane disruption-based intracellular delivery from 1911 until the present. These methods achieve rapid, direct, and universal delivery of almost any cargo molecule or material that can be dispersed in solution. We start by covering the motivations for intracellular delivery and the challenges associated with the different cargo types-small molecules, proteins/peptides, nucleic acids, synthetic nanomaterials, and large cargo. The review then presents a broad comparison of delivery strategies followed by an analysis of membrane disruption mechanisms and the biology of the cell response. We cover mechanical, electrical, thermal, optical, and chemical strategies of membrane disruption with a particular emphasis on their applications and challenges to implementation. Throughout, we highlight specific mechanisms of membrane disruption and suggest areas in need of further experimentation. We hope the concepts discussed in our review inspire scientists and engineers with further ideas to improve intracellular delivery.

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Section: Intracellular Delivery By Direct Penetrationmentioning

confidence: 99%

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

2018

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“…A key challenge for intracellular access is to overcome the physical barrier of the outer membrane of cells without losing cellular functions 4 . Such research efforts have been classified as follows: (1) carrier-based delivery using viral vectors 5 , vesicles 6 , and nanocarriers including lipids 7 and inorganic materials 8 and (2) membrane-disruption delivery via mechanical 9,10 , electrical 11 , thermal 12 , and optical 13 means. Each of these methods has specific advantages and disadvantages in terms of efficiency, invasiveness, safety, dosage control, and cost 4 .…”

Section: Introductionmentioning

confidence: 99%

“…Carbon nanotube- 15 and glass nanopipette-based endoscopes 16 have also provided methods for minimally invasive insertion into cells, and controlled the flow of delivery and extraction of Ca 2+ 17 , proteins 2 , and mRNAs 18 . Examples of this using many nanomaterials have been reported, including vertical Si nanowires 19 , Si nanopillars 20 , Si 3 N 4 11,21 , and Au nanoelectrode 9 . Moreover, aluminum nanostraws 14 , which are arranged in a polycarbonate membrane, have been used as a powerful tool for delivering therapeutic DNA 22 or drugs 23 with improved transfection efficiency.…”

Section: Introductionmentioning

confidence: 99%

Nanostraw membrane stamping for direct delivery of molecules into adhesive cells

Zhang

Miyamoto

et al. 2019

Sci Rep

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Delivering ions and molecules into living cells has become an important challenge in medical and biological fields. Conventional molecular delivery, however, has several issues such as physical and chemical damage to biological cells. Here, we present a method of directly delivering molecules into adhesive cells with an Au-based nanostraw membrane stamp that can physically inject a target molecule into the cytoplasm through a nanostraw duct. We successfully delivered calcein target molecules into adhesive cells with high efficiency (85%) and viability (90%). Furthermore, we modeled the molecular flow through Au nanostraws and then demonstrated the control of calcein flow by changing the concentration and geometry of Au nanostraws. Our Au membrane stamping provides a new way of accessing the cytoplasm to modulate cellular functions via injected molecules.

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“…The generation of genetically engineered animals by introducing gRNA and Cas9 protein into fertilized eggs has already been achieved in many different species of mammals, including ferrets (Yu et al, 2019 ), goats (Ni et al, 2014 ), pigs (Petersen et al, 2016 ) and non-human primates (Kang et al, 2019 ), as well as mice and rats (Shao et al, 2014 ). Recently it has been found that CRISPR complexes can be directly injected into the pronucleus of a fertilized egg using Au nanowire injector (Park et al, 2017 ). This method may improve the efficiency of the generation of transgenic mice by reducing the physical destruction of fertilized eggs.…”

Section: Rapid Generation Of Transgenic Micementioning

confidence: 99%

Methodologies and Challenges for CRISPR/Cas9 Mediated Genome Editing of the Mammalian Brain

2020

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Neurons and glia are highly polarized cells with extensive subcellular structures extending over large distances from their cell bodies. Previous research has revealed elaborate protein signaling complexes localized within intracellular compartments. Thus, exploring the function and the localization of endogenous proteins is vital to understanding the precise molecular mechanisms underlying the synapse, cellular, and circuit function. Recent advances in CRISPR/Cas9-based genome editing techniques have allowed researchers to rapidly develop transgenic animal models and perform single-cell level genome editing in the mammalian brain. Here, we introduce and comprehensively review the latest techniques for genome-editing in whole animals using fertilized eggs and methods for gene editing in specific neuronal populations in the adult or developing mammalian brain. Finally, we describe the advantages and disadvantages of each technique, as well as the challenges that lie ahead to advance the generation of methodologies for genome editing in the brain using the current CRISPR/Cas9 system.

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Suppressing mosaicism by Au nanowire injector-driven direct delivery of plasmids into mouse embryos

Cited by 11 publications

References 75 publications

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

Nanostraw membrane stamping for direct delivery of molecules into adhesive cells

Methodologies and Challenges for CRISPR/Cas9 Mediated Genome Editing of the Mammalian Brain

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