Programmable Inhibition and Detection of RNA Viruses Using Cas13

Molecular Cell, Год журнала: 2019, Номер 76(5), С. 826 - 837.e11

Опубликована: Окт. 10, 2019

Highlights•Thousands of potential Cas13 target sites are present in ssRNA viral genomes•Cas13 is a potent antiviral against three diverse viruses cell culture•Optimal and design criteria identified via genome-wide screen•Companion Cas13-based diagnostics can be used to assess the effects targetingSummaryThe CRISPR effector could an effective for single-stranded RNA (ssRNA) because it programmably cleaves RNAs complementary its (crRNA). Here, we computationally identify thousands crRNA hundreds species that potentially infect humans. We experimentally demonstrate Cas13's activity distinct viruses: lymphocytic choriomeningitis virus (LCMV); influenza A (IAV); vesicular stomatitis (VSV). Combining this with diagnostics, develop Cas13-assisted restriction expression readout (CARVER), end-to-end platform uses detect destroy RNA. further screen crRNAs along LCMV genome evaluate how conservation nucleotide content influence activity. Our results harnessed wide range CARVER's broad utility rapid diagnostic drug development.Graphical abstract

Язык: Английский

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CRISPR/Cas9 editing of endogenous banana streak virus in the B genome of Musa spp. overcomes a major challenge in banana breeding

Communications Biology, Год журнала: 2019, Номер 2(1)

Опубликована: Янв. 31, 2019

Abstract Presence of the integrated endogenous banana streak virus (eBSV) in B genome plantain (AAB) is a major challenge for breeding and dissemination hybrids. As eBSV activates into infectious viral particles under stress, progenitor Musa balbisiana its derivants, having at least one genome, cannot be used as parents crop improvement. Here, we report strategy to inactivate by editing sequences. The regenerated genome-edited events Gonja Manjaya showed mutations targeted sites with potential prevent proper transcription or/and translational functional proteins. Seventy-five percent edited remained asymptomatic comparison non-edited control plants water stress conditions, confirming inactivation particles. This study paves way improvement germplasm use programs produce hybrids that can globally disseminated.

Язык: Английский

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Carrier strategies boost the application of CRISPR/Cas system in gene therapy

Exploration, Год журнала: 2022, Номер 2(2)

Опубликована: Март 15, 2022

Abstract Emerging clustered regularly interspaced short palindromic repeat/associated protein (CRISPR/Cas) genome editing technology shows great potential in gene therapy. However, proteins and nucleic acids suffer from enzymatic degradation the physiological environment low permeability into cells. Exploiting carriers to protect CRISPR system degradation, enhance its targeting of specific tissues cells, reduce immunogenicity is essential stimulate clinical applications. Here, authors review state‐of‐the‐art delivery systems their applications, describe strategies improve safety efficacy mediated editing, categorized by three types cargo formats, that is, Cas: single‐guide RNA ribonucleoprotein, Cas mRNA RNA, plasmid expressing CRISPR/Cas systems. The hope this will help develop safe efficient nanomaterial‐based for tools.

Язык: Английский

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Photoswitchable gRNAs for Spatiotemporally Controlled CRISPR-Cas-Based Genomic Regulation

ACS Central Science, Год журнала: 2020, Номер 6(5), С. 695 - 703

Опубликована: Апрель 1, 2020

The recently discovered CRISPR-Cas gene editing system and its derivatives have found numerous applications in fundamental biology research pharmaceutical sciences. need for precise external control over the regulatory events has driven development of inducible systems. While most light-controllable systems are based on protein engineering, we developed an alternative synthetic approach modification crRNA/tracrRNA duplex (guide RNA or gRNA) with photocaging groups, preventing gRNA from recognizing genome target sequence until deprotection is induced within seconds illumination. This relies a straightforward solid-phase synthesis photocaged gRNAs, simpler purification characterization processes comparison to engineering light-responsive protein. We demonstrated feasibility gRNAs light-mediated DNA cleavage upon brief exposure light vitro. achieved spatiotemporally resolved as well activation cells, whereas showed virtually no detectable absence irradiation. Finally, applied this zebrafish embryos vivo, enabling use strategy developmental tissue applications.

Язык: Английский

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Detect and destroy: CRISPR-based technologies for the response against viruses

Cell Host & Microbe, Год журнала: 2021, Номер 29(5), С. 689 - 703

Опубликована: Апрель 28, 2021

Despite numerous viral outbreaks in the last decade, including a devastating global pandemic, diagnostic and therapeutic technologies remain severely lacking. CRISPR-Cas systems have potential to address these critical needs response against infectious disease. Initially discovered as bacterial adaptive immune system, provide unique opportunity create programmable, sequence-specific for detection of nucleic acids inhibition replication. This review summarizes how systems—in particular recently DNA-targeting Cas12 RNA-targeting Cas13, both possessing trans-cleavage activity—are being harnessed diagnostics therapies. We further highlight whose development has accelerated COVID-19 pandemic.

Язык: Английский

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A CRISPR-Cas Cure for HIV/AIDS

International Journal of Molecular Sciences, Год журнала: 2023, Номер 24(2), С. 1563 - 1563

Опубликована: Янв. 13, 2023

Human immunodeficiency virus (HIV) infections and HIV-induced acquired syndrome (AIDS) continue to represent a global health burden. There is currently no effective vaccine, nor any cure, for HIV infections; existing antiretroviral therapy can suppress viral replication, but only as long antiviral drugs are taken. infects cells of the host immune system, it establish long-lived reservoir, which be targeted edited through gene therapy. Gene editing platforms based on clustered regularly interspaced palindromic repeat-Cas system (CRISPR-Cas) have been recognized promising tools in development therapies infections. In this review, we evaluate current landscape CRISPR-Cas-based against HIV, with an emphasis infection biology well activity restriction factors. We discuss potential combined CRISPR-Cas approach that targets genes activate factors inhibit replication simultaneously. Lastly, focus challenges solutions approaches achieving cure.

Язык: Английский

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CRISPR systems: Novel approaches for detection and combating COVID-19

Virus Research, Год журнала: 2021, Номер 294, С. 198282 - 198282

Опубликована: Янв. 11, 2021

Язык: Английский

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Application of CRISPR/Cas for Diagnosis and Management of Viral Diseases of Banana

Frontiers in Microbiology, Год журнала: 2021, Номер 11

Опубликована: Янв. 27, 2021

Viral diseases are significant biotic constraints for banana ( Musa spp.) production as they affect the yield and limit international movement of germplasm. Among all viruses known to infect banana, bunchy top virus streak widespread economically damaging. The use virus-resistant bananas is most cost-effective option minimize negative impacts viral-diseases on production. CRISPR/Cas-based genome editing emerging powerful tool developing crop varieties in several crops, including banana. availability a vigorous genetic transformation regeneration system well-annotated whole-genome sequence makes it compelling candidate editing. A robust CRISPR/Cas9-based has recently been established, which can be applied disease-resistant varieties. Recently, CRISPR was exploited detect target gene sequences using Cas9, Cas12, Cas13, Cas14 enzymes, thereby unveiling this technology diagnosis. This article presents synopsis recent advancements perspectives application diagnosing resistance against challenges genome-editing

Язык: Английский

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Off-Target Analysis in Gene Editing and Applications for Clinical Translation of CRISPR/Cas9 in HIV-1 Therapy

Frontiers in Genome Editing, Год журнала: 2021, Номер 3

Опубликована: Авг. 17, 2021

As genome-editing nucleases move toward broader clinical applications, the need to define limits of their specificity and efficiency increases. A variety approaches for nuclease cleavage detection have been developed, allowing a full-genome survey targeting landscape repair outcomes nuclease-induced double-strand breaks. Each approach has advantages disadvantages relating means target-site capture, target enrichment mechanism, cellular environment, false discovery, validation bona fide off-target sites in cells. This review examines strengths, limitations, origins different classes systems including anchored primer (GUIDE-seq), situ (BLISS), vitro selection libraries (CIRCLE-seq), chromatin immunoprecipitation (ChIP) (DISCOVER-Seq), translocation sequencing (LAM PCR HTGTS), genomic DNA digestion (Digenome-seq SITE-Seq). Emphasis is placed on specific modifications that give rise enhanced performance contemporary techniques over predecessors comparative applications. The relevance these discussed context assessing safety novel CRISPR/Cas9 HIV-1 curative strategies. With recent success SIV-1 viral suppression humanized mice non-human primates, respectively, using CRISPR/Cas9, rigorous exploration potential effects critical importance. Such analyses would benefit from application this review.

Язык: Английский

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A histone deacetylase network regulates epigenetic reprogramming and viral silencing in HIV-infected cells

Cell chemical biology, Год журнала: 2023, Номер 30(12), С. 1617 - 1633.e9

Опубликована: Дек. 1, 2023

Язык: Английский

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