Magnetically guided non-invasive CRISPR-Cas9/gRNA delivery across blood-brain barrier to eradicate latent HIV-1 infection

Scientific Reports, Journal Year: 2019, Volume and Issue: 9(1)

Published: March 8, 2019

Abstract CRISPR-Cas9/gRNA exhibits therapeutic efficacy against latent human immunodeficiency virus (HIV) genome but the delivery of this cargo to brain remains as a challenge. In research, for first time, we demonstrated magnetically guided non-invasive nano-formulation (NF), composed Cas9/gRNA bound with magneto-electric nanoparticles (MENPs), across blood-brain barrier (BBB) inhibit HIV-1 infection in microglial (hμglia)/HIV (HC69) cells. An optimized ac-magnetic field 60 Oe was applied on NF release from MENPs surface and facilitate cell uptake resulting intracellular inhibition HIV. The outcomes suggested that developed reduced HIV-LTR expression significantly comparison unbound HIV hμglia/HIV These findings were also validated qualitatively using fluorescence microscopy assess microglia We believe CNS (CRISPR/Cas9-gRNA-MENPs) BBB certainly will have clinical utility future personalized nanomedicine manage neuroHIV/AIDS.

Language: Английский

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Advances in therapeutic CRISPR/Cas9 genome editing

Translational research, Journal Year: 2015, Volume and Issue: 168, P. 15 - 21

Published: Sept. 26, 2015

Targeted nucleases are widely used as tools for genome editing. Two years ago the clustered regularly interspaced short palindromic repeat (CRISPR)-associated Cas9 nuclease was first time, and since then has largely revolutionized field. The tremendous success of CRISPR/Cas9 editing tool is powered by ease design principle guide RNA that targets to desired DNA locus, high specificity efficiency CRISPR/Cas9-generated breaks. Several studies recently successfully modulate disease-causing alleles in vivo animal models ex somatic induced pluripotent stem cells, raising hope therapeutic clinics. In this review, we will summarize discuss such preclinical gene therapy reports.

Language: Английский

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CRISPR/Cas9-Derived Mutations Both Inhibit HIV-1 Replication and Accelerate Viral Escape

Cell Reports, Journal Year: 2016, Volume and Issue: 15(3), P. 481 - 489

Published: April 1, 2016

Cas9 cleaves specific DNA sequences with the assistance of a programmable single guide RNA (sgRNA). Repairing this broken by cell's error-prone non-homologous end joining (NHEJ) machinery leads to insertions and deletions (indels) that often impair function. Using HIV-1, we have now demonstrated many these indels are indeed lethal for virus, but others lead emergence replication competent viruses resistant Cas9/sgRNA. This unexpected contribution development viral resistance is facilitated some not deleterious replication, refractory recognition same sgRNA as result changing target sequences. observation illustrates two opposite outcomes Cas9/sgRNA action, i.e., inactivation HIV-1 acceleration escape, thereby potentially limiting use in therapy.

Language: Английский

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Monocytes and Macrophages as Viral Targets and Reservoirs

International Journal of Molecular Sciences, Journal Year: 2018, Volume and Issue: 19(9), P. 2821 - 2821

Published: Sept. 18, 2018

Viruses manipulate cell biology to utilize monocytes/macrophages as vessels for dissemination, long-term persistence within tissues and virus replication. enter cells through endocytosis, phagocytosis, macropinocytosis or membrane fusion. These processes play important roles in the mechanisms contributing pathogenesis of these agents establishing viral genome latency. Upon infection, monocytes respond with an elevated expression proinflammatory signalling molecules antiviral responses, is shown case influenza, Chikungunya, human herpes Zika viruses. Human immunodeficiency initiates acute inflammation on site during early stages infection but there a shift M1 M2 at later infection. Cytomegalovirus creates balance between pro- anti-inflammatory by inducing specific phenotype M1/M2 continuum. Despite facilitating inflammation, infected macrophages generally display abolished apoptosis restricted cytopathic effect, which sustains production. The majority viruses discussed this review employ repository certain use productive This focuses adaptations monocytes/macrophages, immune escape, reprogramming response host cells.

Language: Английский

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Generation of Blastocyst-like Structures from Mouse Embryonic and Adult Cell Cultures

Cell, Journal Year: 2019, Volume and Issue: 179(3), P. 687 - 702.e18

Published: Oct. 1, 2019

Language: Английский

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CRISPR-Cas9 Can Inhibit HIV-1 Replication but NHEJ Repair Facilitates Virus Escape

Molecular Therapy, Journal Year: 2016, Volume and Issue: 24(3), P. 522 - 526

Published: Jan. 22, 2016

Several recent studies demonstrated that the clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease Cas9 can be used for guide RNA (gRNA)-directed, sequence-specific cleavage of HIV proviral DNA in infected cells. We here demonstrate profound inhibition HIV-1 replication by harnessing T cells with and antiviral gRNAs. However, virus rapidly consistently escaped from this inhibition. Sequencing escape variants revealed nucleotide insertions, deletions, substitutions around Cas9/gRNA site are typical repair nonhomologous end-joining pathway. thus potency CRISPR-Cas9 as an approach, but any therapeutic strategy should consider viral implications.

Language: Английский

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Excision of HIV-1 DNA by gene editing: a proof-of-concept in vivo study

Gene Therapy, Journal Year: 2016, Volume and Issue: 23(8-9), P. 690 - 695

Published: May 19, 2016

Language: Английский

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Next-Generation Sequencing — An Overview of the History, Tools, and “Omic” Applications

InTech eBooks, Journal Year: 2016, Volume and Issue: unknown

Published: Jan. 14, 2016

Next-generation sequencing (NGS) technologies using DNA, RNA, or methylation have impacted enormously on the life sciences. NGS is choice for large-scale genomic and transcriptomic because of high-throughput production outputs data in gigabase range per instrument run lower cost compared to traditional Sanger first-generation method. The vast amounts generated by broadened our understanding structural functional genomics through concepts “omics” ranging from basic integrated systeomics, providing new insight into workings meaning genetic conservation diversity living things. today more than ever about how different organisms use information molecular biology survive reproduce with without mutations, disease, within their population networks changing environments. In this chapter, advances, applications, challenges are reviewed starting a history followed major platforms, bioinformatics issues confronting storage analysis, impacts made fields genetics, biology, agriculture, medicine brave, world ”omics.”

Language: Английский

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Application of CRISPR/Cas9-Based Gene Editing in HIV-1/AIDS Therapy

Frontiers in Cellular and Infection Microbiology, Journal Year: 2019, Volume and Issue: 9

Published: March 22, 2019

Despite the fact that great efforts have been made in prevention and resistance of HIV-1 infection, HIV-1/AIDS remains a major threat to global human health. Highly active antiretroviral therapy (HAART) can suppress virus replication, but it cannot eradicate latent viral reservoirs patients. Recently, clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system has engineered as an effective gene-editing technology with potential treat HIV-1/AIDS. It be used target cellular co-factors or genome reduce infection clear provirus, well induce transcriptional activation for elimination. This versatile gene editing successfully applied reduction cells animal models. Here, we update rapid progress CRISPR/Cas9-based research recent years discuss limitations future perspectives its application.

Language: Английский

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Block-And-Lock Strategies to Cure HIV Infection

Viruses, Journal Year: 2020, Volume and Issue: 12(1), P. 84 - 84

Published: Jan. 10, 2020

Today HIV infection cannot be cured due to the presence of a reservoir latently infected cells inducing viral rebound upon treatment interruption. Hence, latent is considered as major barrier for an cure. So far, efforts completely eradicate via shock-and-kill approach have proven difficult and unsuccessful. Therefore, more research has been done recently on alternative block-and-lock functional cure strategy. In contrast strategy that aims entire reservoir, permanently silence all proviruses, even after silencing can achieved by targeting different factors transcription machinery. this review, we first describe underlying mechanisms silencing. Next, give overview strategies under investigation.

Language: Английский

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