Evidence map›Paper›PMID 37292682›Full record

ArticlebioRxiv : the preprint server for biology2023

Rapid cloning-free mutagenesis of new SARS-CoV-2 variants using a novel reverse genetics platform.

Enja Kipfer, David Hauser, Martin J Lett, Fabian Otte, Lorena Urda, Yuepeng Zhang, Christopher M R Lang, Mohamed Chami, Christian Mittelholzer, Thomas Klimkait

Abstract readPreprint
PubMed Full text at PMC Publisher
In one paragraph

Article in bioRxiv : the preprint server for biology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

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.

2 · The registry

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.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

10 authors.

Enja KipferMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.
David HauserMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.
Martin J LettMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.
Fabian OtteMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.
Lorena UrdaMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.
Yuepeng ZhangMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.
Christopher M R LangMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.
Mohamed ChamiBioEM Lab, Biozentrum, University of Basel, Mattenstrasse 26, 4058 Basel, Switzerland.
Christian MittelholzerMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.
Thomas KlimkaitMolecular Virology, Department of Biomedicine, University of Basel, Petersplatz 10, 4009 Basel, Switzerland.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Reverse genetic systems enable the engineering of RNA virus genomes and are instrumental in studying RNA virus biology. With the recent outbreak of the COVID-19 pandemic, already established methods were challenged by the large genome of SARS-CoV-2. Herein we present an elaborated strategy for the rapid and straightforward rescue of recombinant plus-stranded RNA viruses with high sequence fidelity, using the example of SARS-CoV-2. The strategy called CLEVER (CLoning-free and Exchangeable system for Virus Engineering and Rescue) is based on the intracellular recombination of transfected overlapping DNA fragments allowing the direct mutagenesis within the initial PCR-amplification step. Furthermore, by introducing a linker fragment - harboring all heterologous sequences - viral RNA can directly serve as a template for manipulating and rescuing recombinant mutant virus, without any cloning step. Overall, this strategy will facilitate recombinant SARS-CoV-2 rescue and accelerate its manipulation. Using our protocol, newly emerging variants can quickly be engineered to further elucidate their biology. To demonstrate its potential as a reverse genetics platform for plus-stranded RNA viruses, the protocol has been successfully applied for the cloning-free rescue of recombinant Chikungunya and Dengue virus.

Identifiers

PMID37292682
PMCPMC10245781

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Registered trials

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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.