Abstract:
:A facile and efficient method for the precise editing of large viral genomes is required for the selection of attenuated vaccine strains and the construction of gene therapy vectors. The type II prokaryotic CRISPR-Cas (clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas)) RNA-guided nuclease system can be introduced into host cells during viral replication. The CRISPR-Cas9 system robustly stimulates targeted double-stranded breaks in the genomes of DNA viruses, where the non-homologous end joining (NHEJ) and homology-directed repair (HDR) pathways can be exploited to introduce site-specific indels or insert heterologous genes with high frequency. Furthermore, CRISPR-Cas9 can specifically inhibit the replication of the original virus, thereby significantly increasing the abundance of the recombinant virus among progeny virus. As a result, purified recombinant virus can be obtained with only a single round of selection. In this study, we used recombinant adenovirus and type I herpes simplex virus as examples to demonstrate that the CRISPR-Cas9 system is a valuable tool for editing the genomes of large DNA viruses.
journal_name
PLoS Pathogjournal_title
PLoS pathogensauthors
Bi Y,Sun L,Gao D,Ding C,Li Z,Li Y,Cun W,Li Qdoi
10.1371/journal.ppat.1004090subject
Has Abstractpub_date
2014-05-01 00:00:00pages
e1004090issue
5eissn
1553-7366issn
1553-7374pii
PPATHOGENS-D-13-02499journal_volume
10pub_type
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