PRJNA564604
Effects of anionic polymer-stabilized Cas9 RNPs on off-target genome modifications at EMX1
The anionic polymers PGA or ssODNenh can increase NHEJ editing efficiency, including at off-target sites. CD3+ T cells isolated from two healthy donors were treated with regular, PGA-stabilized, or ssODNenh-stabilized RNPs incorporating a gRNA targeting the EMX1 gene previously identified by CIRCLE-Seq to create multiple off-target double-strand breaks. Control cells were electroporated without RNP. On- and off-target genomic regions were PCRamplified, and editing outcomes were quantified by deep sequencing on an Illumina MiniSeq sequencer.