<u>Ad</u>vanced <u>v</u>iral genome <i><u>i</u>n vitro</i> <u>C</u>as9 <u>e</u>diting (AdVICE): an overnight method for traceless and limitless manipulation of adenoviral and vector genomes with large transgenes.

ABSTRACT: The size and complexity of large viral genomes limit the technical possibilities for genome manipulations in fundamental research and medical or technological applications. State-of-the-art recombineering in bacteria has partially overcome this limit but remains a time-consuming and complex procedure requiring specialist expertise. Here, we describe a simplified and highly efficient in vitro protocol for unlimited and traceless manipulation applicable to large viral genomes from DNA viruses using a combination of CRISPR/Cas9 cleavage and in vitro DNA assembly. We successfully used the protocol to manipulate adenovirus genomes, showing that genome rescue from viruses, insertions, deletions, and mutagenesis can be performed in a simple overnight procedure in a standard laborato

SUBMITTER: Vergnes J-B 

PROVIDER: S-EPMC12172474 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature