Genome-wide knockout libraries for entire classes of genomic elements in humans and other species.

Abstract

(A) An example of paired sgRNAs designed against the upstream ultraconserved element (UCE) and promoter of the human IRX3 gene. IRX3 lies on the antisense strand. The exact target regions are shown in black, flanked by the design regions in green. The ten sgRNA pairs for each are denoted by red bars. Integrated chromatin marks from the ENCODE project [26] are displayed below, in addition to PhyloP multispecies conservation scores [33]. Note the region of elevated conservation corresponding to the UCE. (B,C) Summary of paired sgRNA designs targeting entire classes of genomic elements. In each figure, the left scale and grey bars represent the design performance, as in Fig 2. The right scale and black bars indicate the total number of elements in each class. (B) shows a series of genomic element classes for human, while (C) shows designs for the entire set of annotated microRNA genes in five species. Designs were created with default settings; designs using “DECKO” construction method give identical results.</p