CRISPR-Csy4-Mediated Editing of Rotavirus Double-Stranded RNA Genome

TY - JOUR

T1 - CRISPR-Csy4-Mediated Editing of Rotavirus Double-Stranded RNA Genome

AU - Papa, Guido

AU - Venditti, Luca

AU - Braga, Luca

AU - Schneider, Edoardo

AU - Giacca, Mauro

AU - Petris, Gianluca

AU - Burrone, Oscar R.

PY - 2020/9/29

Y1 - 2020/9/29

N2 - CRISPR-nucleases have been widely applied for editing cellular and viral genomes, but nuclease-mediated genome editing of double-stranded RNA (dsRNA) viruses has not yet been reported. Here, by engineering CRISPR-Csy4 nuclease to localize to rotavirus viral factories, we achieve the nuclease-mediated genome editing of rotavirus, an important human and livestock pathogen with a multisegmented dsRNA genome. Rotavirus replication intermediates cleaved by Csy4 is edited through the formation of precise deletions in the targeted genome segments in a single replication cycle. Using CRISPR-Csy4-mediated editing of rotavirus genome, we label the products of rotavirus secondary transcription made by newly assembled viral particles during rotavirus replication, demonstrating that this step largely contributes to the overall production of viral proteins. We anticipate that the nuclease-mediated cleavage of dsRNA virus genomes will promote an advanced level of understanding of viral replication and host-pathogen interactions, also offering opportunities to develop therapeutics.

AB - CRISPR-nucleases have been widely applied for editing cellular and viral genomes, but nuclease-mediated genome editing of double-stranded RNA (dsRNA) viruses has not yet been reported. Here, by engineering CRISPR-Csy4 nuclease to localize to rotavirus viral factories, we achieve the nuclease-mediated genome editing of rotavirus, an important human and livestock pathogen with a multisegmented dsRNA genome. Rotavirus replication intermediates cleaved by Csy4 is edited through the formation of precise deletions in the targeted genome segments in a single replication cycle. Using CRISPR-Csy4-mediated editing of rotavirus genome, we label the products of rotavirus secondary transcription made by newly assembled viral particles during rotavirus replication, demonstrating that this step largely contributes to the overall production of viral proteins. We anticipate that the nuclease-mediated cleavage of dsRNA virus genomes will promote an advanced level of understanding of viral replication and host-pathogen interactions, also offering opportunities to develop therapeutics.

KW - CRISPR

KW - Csy4

KW - deletions

KW - dsRNA

KW - genome editing

KW - NSP5

KW - repair

KW - rotavirus

KW - secondary transcription

KW - viroplasm

UR - http://www.scopus.com/inward/record.url?scp=85091790427&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2020.108205

DO - 10.1016/j.celrep.2020.108205

M3 - Article

AN - SCOPUS:85091790427

SN - 2211-1247

VL - 32

JO - Cell Reports

JF - Cell Reports

IS - 13

M1 - 108205

ER -