High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases

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The clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) system was discovered as a component of the bacterial acquired immune system that cleaves foreign DNA. This system is now used for site-specific genome editing in a wide range of organisms, including bacteria, yeasts, plants, and animals. However, the use of this approach in non-cell organisms, such as non-integrating viruses, has not been reported. Because multiple steps are required to construct mutant or recombinant DNA viruses with large genomes using the current approaches, we used the CRISPR-Cas9 system to introduce site-specific indels and insert a foreign gene into an adenoviral vector and wild-type herpes simplex virus. The high efficiency of CRISPR-Cas9 editing allowed for simple construction and purification of recombinant progeny virus. We believe that this new technique will have broad practical significance for selecting attenuated vaccine strains and antiviral drugs, constructing gene therapy vectors, and establishing efficient methods for viral biological studies.

Vyšlo v časopise: High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases. PLoS Pathog 10(5): e32767. doi:10.1371/journal.ppat.1004090
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004090

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