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 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 direc

SUBMITTER: Kipfer E 

PROVIDER: S-EPMC10245781 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature