NextSeq 500 sequencing; Genome-scale CRISPR-Cas9 knockout screens for enterovirus D68 and rhinovirus C15
Genome-scale CRISPR-Cas9 knockout screens for enterovirus D68 and rhinovirus C15
>1000-fold coverage of mutagenized cells (libraries A and B) was infected with either RV-C15 (MOI=1 PFU/cell) or EV-D68 Missouri (MOI=1 PFU/cell).  RV-C15 infection was repeated for an additional round at 6 days post-infection. As soon as appearance of visibly viable colonies was observed, populations of virus-resistant cells were pooled and harvested. Uninfected starting populations of mutagenized cells were used as the unselected reference.  Total genomic DNA from both virus-resistant and uninfected cells was respectively extracted using QIAamp DNA Mini Kit (Qiagen).  The inserted guide RNA sequences were retrieved from the genomic DNA by PCR amplification.
Experimental Factor: infect
EV-D68 (Missouri) infected