ArticleMethods in molecular biology (Clifton, N.J.)2017
Efficient Reverse Genetic Systems for Rapid Genetic Manipulation of Emergent and Preemergent Infectious Coronaviruses.
Article in Methods in molecular biology (Clifton, N.J.), 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
15 citing papers in PubMed.
- A low-background, fluorescent assay to evaluate inhibitors of diverse viral proteases.Journal of virology · 2023Article
- An in vitro workflow to create and modify infectious clones using replication cycle reaction.Virology · 2023Article
- Molecular and Genetics-Based Systems for Tracing the Evolution and Exploring the Mechanisms of Human Norovirus Infections.International journal of molecular sciences · 2023Review
- Assembly of plant virus agroinfectious clones using biological material or DNA synthesis.STAR protocols · 2022Article
- Engineering and Characterization of Avian Coronavirus Mutants Expressing Fluorescent Reporter Proteins from the Replicase Gene.Journal of virology · 2022Article
- The repeated setbacks of HIV vaccine development laid the groundwork for SARS-CoV-2 vaccines.Health policy and technology · 2022Article
- Recovery Infectious Enterovirus 71 by Bac-to-Bac Expression SystemFrontiers in microbiology · 2022Article
- An appeal for an objective, open, and transparent scientific debate about the origin of SARS-CoV-2.Lancet (London, England) · 2021Article
- A versatile reverse genetics platform for SARS-CoV-2 and other positive-strand RNA viruses.Nature communications · 2021Article
- Immunity, virus evolution, and effectiveness of SARS-CoV-2 vaccines.Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologica · 2021Review
- Review
- Reverse genetic systems: Rational design of coronavirus live attenuated vaccines with immune sequelae.Advances in virus research · 2020Review
- Genetically Engineering a Susceptible Mouse Model for MERS-CoV-Induced Acute Respiratory Distress Syndrome.Methods in molecular biology (Clifton, N.J.) · 2020Article
- Broad-spectrum coronavirus antiviral drug discovery.Expert opinion on drug discovery · 2019Review
- Modeling pathogenesis of emergent and pre-emergent human coronaviruses in mice.Mammalian genome : official journal of the International Mammalian Genome Society · 2018Review
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Emergent and preemergent coronaviruses (CoVs) pose a global threat that requires immediate intervention. Rapid intervention necessitates the capacity to generate, grow, and genetically manipulate infectious CoVs in order to rapidly evaluate pathogenic mechanisms, host and tissue permissibility, and candidate antiviral therapeutic efficacy. CoVs encode the largest viral RNA genomes at about 28-32,000 nucleotides in length, and thereby complicate efficient engineering of the genome. Deconstructing the genome into manageable fragments affords the plasticity necessary to rapidly introduce targeted genetic changes in parallel and assort mutated fragments while maximizing genome stability over time. In this protocol we describe a well-developed reverse genetic platform strategy for CoVs that is comprised of partitioning the viral genome into 5-7 independent DNA fragments (depending on the CoV genome), each subcloned into a plasmid for increased stability and ease of genetic manipulation and amplification. Coronavirus genomes are conveniently partitioned by introducing type IIS or IIG restriction enzyme recognition sites that confer directional cloning. Since each restriction site leaves a unique overhang between adjoining fragments, reconstruction of the full-length genome can be achieved through a standard DNA ligation comprised of equal molar ratios of each fragment. Using this method, recombinant CoVs can be rapidly generated and used to investigate host range, gene function, pathogenesis, and candidate therapeutics for emerging and preemergent CoVs both in vitro and in vivo.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.