Fig. 1

Strategy for Cpf1 multiplex CRISPR editing. ASchematic depiction of gene knock-out with a gRNA redundancy strategy. Multiple gRNAs are used to induce a double stranded break. Homology-directed repair using a dsDNA gene targeting substrate (GTS) as template induces a 300 bp deletion at the start of the gene.BSchematic for USER assembly of a Cpf1 CRISPR vector containing one gRNA using two USER compatible PCR fragments.CSchematic depiction of expression of a Cpf1 multi-guide precursor RNA from a MEC controlled by a U3 promoter and terminator, followed by processing of the precursor RNA by Cpf1, RNase Z and tRNase P to yield mature gRNAs.DSchematic USER assembly of a 9-gRNA Cpf1 MEC using two USER compatible PCR fragments and five gRNA bio-blocks. Graphics legend is presented in the box