Читать книгу Principles of Virology - Jane Flint, S. Jane Flint - Страница 152

Mutations can be introduced into a viral genome when it is cloned in its entirety. Mutagenesis is usually carried out on cloned subfragments, which are then substituted into full-length cloned DNA. This step can now be bypassed by using CRISPR/Cas9 to introduce mutations into complete DNA copies of viral genomes. Viruses are then recovered by introduction of the mutagenized DNA into cultured cells by transfection. This approach has been applied to cloned DNA copies of RNA and DNA viral genomes.

Introduction of mutagenized viral nucleic acid into cultured cells by transfection may have a variety of outcomes, ranging from no effect to a complete block of viral reproduction. Whether the introduced mutation is responsible for an observed phenotype deserves careful scrutiny (Box 3.10).