This efficiency, coupled with the simplicity of constructing guide RNAs from synthetic oligonucleotides, which are amenable to being produced in both multi-well plates or in larger pools of tens of thousands, has allowed both arrayed (Hultquist et al. 2016) and pooled screening strategies (Gilbert et al. 2014; Koike-Yusa et al. 2014; Konermann et al. 2015; Shalem et al. 2014; Wang et al. 2014a, b) to be conceived. This provides the exciting prospect of forward genetic screening in human patient-derived cell lines to identify potential therapeutic targets and to better understand molecular and genetic basis of disease (Fig. 1).
