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

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 coronavirus disease 2019 pandemic, already established methods were challenged by the large genome of severe acute respiratory syndrome coronavirus 2 (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 frag

SUBMITTER: Kipfer ET 

PROVIDER: S-EPMC10662946 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature