Editorial focus: understanding off-target effects as the key to successful RNAi therapy

Bartoszewski, Rafał; Sikorski, Aleksander F.

doi:10.1186/s11658-019-0196-3

Cited by 95 publications

(74 citation statements)

References 396 publications

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“…Hence, the hypothesis that the lung cells lack specific miRNA directed against pathogenic HCoVs seems rather unlikely, although it still cannot be excluded for specific miRNAs. This observation questions the use of miRNA overexpression (miRNA mimics) as a therapeutic approach against the COVID-19 infection since siRNAs directed against viral RNA would be more specific and therefore less prone to have off-target effects ( 12 ).…”

Section: Comparison Of the Mirna Target Sites In Human Coronavirusesmentioning

confidence: 99%

SARS-CoV-2 may regulate cellular responses through depletion of specific host miRNAs

Bartoszewski

Dąbrowski

Jakieła

et al. 2020

American Journal of Physiology-Lung Cellular and Molecular Phys

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Cold viruses have generally been considered fairly innocuous until the appearance of the severe acute respiratory coronavirus 2 (SARS-CoV-2) in 2019 which caused the coronavirus disease 2019 (COVID-19) global pandemic. Two previous viruses foreshadowed that a coronavirus could potentially have devastating consequences in 2002 (severe acute respiratory coronavirus (SARS-CoV)) and in 2012 (middle east respiratory syndrome coronavirus (MERS-CoV)). The question that arises is why these viruses are so different from the relatively harmless cold viruses. Based on an analysis of the current literature and using bioinformatic approaches, we examined the potential human miRNA interactions with the SARS-CoV-2's genome, and compared the miRNA target sites in seven coronavirus genomes that include SARS-CoV-2, MERS-CoV, SARS-CoV, and four non-pathogenic coronaviruses. Here, we discuss the possibility that pathogenic HCoVs including SARS-CoV-2 could modulate host miRNA levels by acting as miRNA sponges to facilitate viral replication and/or to avoid immune responses.

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Section: Comparison Of the Mirna Target Sites In Human Coronavirusesmentioning

confidence: 99%

SARS-CoV-2 may regulate cellular responses through depletion of specific host miRNAs

Bartoszewski

Dąbrowski

Jakieła

et al. 2020

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Then, the pNc-SAG1::CAS9-U6::sgGRA6 plasmid and selectable marker of DHFR-TS were electroporated into the Nc-1 strain, which was inoculated into HFF cells and allowed screening under pyrimethamine selection pressure. It is worth mentioning that gene knockout using stable transfection can avoid the common off-target effects brought by transient transfection using the RNAi method (33). In addition, the gene knockout method can completely remove the function of GRA6; however, the RNAi method can only lower the function of GRA6 to some extent, which may influence the following function research of GRA6.…”

Section: Discussionmentioning

confidence: 99%

Effects of Dense Granular Protein 6 (GRA6) Disruption on Neospora caninum Virulence

et al. 2020

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Neospora caninum (N. caninum) is a major cause of abortions in cattle. During its invasion of host cells, a parasitophorous vacuole (PV) is formed, accompanied by an intravacuolar network (IVN). The IVN takes part in parasite ingesting of nutrients from hosts. The dense granular proteins of N. caninum (NcGRAs) play a key role in forming the PV and the IVN, which may influence virulence during N. caninum invasion. The present study aimed to explore the biological function of NcGRA6 in N. caninum by disrupting the NcGRA6 gene in the Nc-1 strain. We successfully constructed an NcGRA6-targeting CRISPR plasmid (pNc-SAG1-Cas9:U6-SgGRA6) and amplified the DHFR-TS DNA donor. The NcGRA6 knockout mutation (NcGRA6) was generated by co-electroporation of the pNc-SAG1::CAS9-U6::sgGRA6 plasmid and the DHFR-TS DNA donor into the Nc-1 strain, which was then cultured under pyrimethamine selection pressure. The NcGRA6 mutation was further verified by identification of NcGRA6 gene disruption using PCR, measurement of NcGRA6 gene transcription levels using qPCR, assessment of NcGRA6 protein expression levels using western blotting, and observation of NcGRA6 protein location using immunofluorescence and immunoelectron microscopy. The results of in vitro virulence assays, including plaque, invasion, egress, and replication assays, showed that the NcGRA6 strain had smaller plaques, similar invasion and egress ability, and slower intracellular replication ability than the Nc-1 strain. The results of in vivo virulence assays showed that the NcGRA6 strain exhibited reduced virulence and improved survival ability in mice compared with the Nc-1 strain. The parasite burden in NcGRA6 strain-infected mouse tissues, including the heart, brain, liver, spleen, lung, and kidney, was significantly reduced compared with that in mice infected with the Nc-1 strain. These data suggest that we successfully constructed a NcGRA6 strain and verify that NcGRA6 is a critical virulence factor. NcGRA6 gene disruption can slow down N. caninum proliferation and lower the pathogenicity to hosts. Our findings provide a foundation for future research on other targeted N. caninum protein functions and may help in exploring the interaction mechanisms between parasites and hosts.

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“…In addition, chemically modified double-stranded miRNAs of miRNA-23b and miRNA-25 were shown to inhibit podocyte injury in diabetic nephropathy mouse models in vivo (Zhao et al, 2016;Liu et al, 2017). However, there are still unresolved aspects of treatment of podocyte injury with miRNAs with/without vectors, such as efficient cellular uptake and intracellular re-release, as well as targeted delivery (Bartoszewski and Sikorski, 2019).…”

Section: Methods Of Modulation Of Mirna Expression Levels On Podocytementioning

confidence: 99%

MicroRNAs in Podocyte Injury in Diabetic Nephropathy

et al. 2020

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Diabetic nephropathy is one of the major complications of diabetes mellitus and is the leading cause of end-stage renal disease worldwide. Podocyte injury contributes to the development of diabetic nephropathy. However, the molecules that regulate podocyte injury in diabetic nephropathy have not been fully clarified. MicroRNAs (miRNAs) are small non-coding RNAs that can inhibit the translation of target messenger RNAs. Previous reports have described alteration of the expression levels of many miRNAs in cultured podocyte cells stimulated with a high glucose concentration and podocytes in rodent models of diabetic nephropathy. The associations between podocyte injury and miRNA expression levels in blood, urine, and kidney in patients with diabetic nephropathy have also been reported. Moreover, modulation of the expression of several miRNAs has been shown to have protective effects against podocyte injury in diabetic nephropathy in cultured podocyte cells in vitro and in rodent models of diabetic nephropathy in vivo. Therefore, this review focuses on miRNAs in podocyte injury in diabetic nephropathy, with regard to their potential as biomarkers and miRNA modulation as a therapeutic option.

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Editorial focus: understanding off-target effects as the key to successful RNAi therapy

Cited by 95 publications

References 396 publications

SARS-CoV-2 may regulate cellular responses through depletion of specific host miRNAs

SARS-CoV-2 may regulate cellular responses through depletion of specific host miRNAs

Effects of Dense Granular Protein 6 (GRA6) Disruption on Neospora caninum Virulence

MicroRNAs in Podocyte Injury in Diabetic Nephropathy

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