Get out or die trying: Peptide- and protein-based endosomal escape of RNA therapeutics

Klipp, A.; Burger, Michael; Leroux, Jean‐Christophe

doi:10.1016/j.addr.2023.115047

“…Generally, transfection agents require affinity to the cellular membrane, glycocalyx, or specific receptors to mediate endocytosis [ 26 , 86 – 89 ]. The stronger the interactions, however, the more difficult it is to remove non-endocytosed DNA from the cellular membrane during sample preparation.…”

Section: Covalently Labeled Dnamentioning

confidence: 99%

The intracellular visualization of exogenous DNA in fluorescence microscopy

Greitens,

Leroux,

Burger

2024

Drug Deliv. and Transl. Res.

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In the development of non-viral gene delivery vectors, it is essential to reliably localize and quantify transfected DNA inside the cell. To track DNA, fluorescence microscopy methods are commonly applied. These mostly rely on fluorescently labeled DNA, DNA binding proteins fused to a fluorescent protein, or fluorescence in situ hybridization (FISH). In addition, co-stainings are often used to determine the colocalization of the DNA in specific cellular compartments, such as the endolysosomes or the nucleus. We provide an overview of these DNA tracking methods, advice on how they should be combined, and indicate which co-stainings or additional methods are required to draw precise conclusions from a DNA tracking experiment. Some emphasis is given to the localization of exogenous DNA inside the nucleus, which is the last step of DNA delivery. We argue that suitable tools which allow for the nuclear detection of faint signals are still missing, hampering the rational development of more efficient non-viral transfection systems. Graphical abstract The intracellular visualization of exogenous DNA in fluorescence microscopy. Created with BioRender.com.

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“…Generally, transfection agents require affinity to the cellular membrane, glycocalyx, or specific receptors to mediate endocytosis [16,[77][78][79][80]. The stronger the interactions, however, the more difficult it is to remove non-endocytosed DNA from the cellular membrane during sample preparation.…”

Section: Cellular Uptakementioning

confidence: 99%

The intracellular visualization of exogenous DNA in fluorescence microscopy

Greitens,

Leroux,

Burger

2023

Preprint

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In the development of non-viral gene delivery vectors, it is essential to reliably localize and quantify transfected DNA inside the cell. To track DNA, fluorescence microscopy methods are commonly applied. These mostly rely on fluorescently labeled DNA, DNA binding proteins fused to a fluorescent protein, or fluorescence in situ hybridization (FISH). In addition, co-stainings are often used to determine the colocalization of the DNA in specific cellular compartments, such as the endolysosomes or the nucleus. We provide an overview of these DNA tracking methods, advice on how they should be combined, and indicate which co-stainings or additional methods are required to draw precise conclusions from a DNA tracking experiment. Some emphasis is given to the localization of exogenous DNA inside the nucleus, which is the last step of DNA delivery. We argue that suitable tools which allow for the nuclear detection of faint signals are still missing, hampering the rational development of more efficient non-viral transfection systems.

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“…Non-coding RNA (ncRNA) refers to RNA that does not encode proteins [57,58]. Only 1-2% of the human genome encodes proteins, but up to 90% of the transcripts produced by the genome have no protein-coding ability.…”

Section: Non-coding Rnas (Ncrnas)mentioning

confidence: 99%

Insight into Steroid-Induced ONFH: The Molecular Mechanism and Function of Epigenetic Modification in Mesenchymal Stem Cells

Huang,

Qing,

Xiao

et al. 2023

Biomolecules

4

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Osteonecrosis of the femoral head (ONFH) is a common refractory orthopedic disease, which is one of the common causes of hip pain and dysfunction. ONFH has a very high disability rate, which is associated with a heavy burden to patients, families, and society. The pathogenesis of ONFH is not completely clear. At present, it is believed that it mainly includes coagulation dysfunction, abnormal lipid metabolism, an imbalance of osteogenic/adipogenic differentiation, and poor vascularization repair. The prevention and treatment of ONFH has always been a great challenge for clinical orthopedic surgeons. However, recent studies have emphasized that the use of mesenchymal stem cells (MSCs) to treat steroid-induced ONFH (SONFH) is a promising therapy. This review focuses on the role and molecular mechanism of epigenetic regulation in the progress of MSCs in the treatment of SONFH, and discusses the significance of the latest research in the treatment of SONFH from the perspective of epigenetics.

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Get out or die trying: Peptide- and protein-based endosomal escape of RNA therapeutics

Cited by 23 publications

References 153 publications

The intracellular visualization of exogenous DNA in fluorescence microscopy

The intracellular visualization of exogenous DNA in fluorescence microscopy

The intracellular visualization of exogenous DNA in fluorescence microscopy

Insight into Steroid-Induced ONFH: The Molecular Mechanism and Function of Epigenetic Modification in Mesenchymal Stem Cells

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