Skip to main content
. 2020 Jun 18;41(11):1427–1432. doi: 10.1038/s41401-020-0452-0

Fig. 2. Design of plasmid containing eSpCas9(1.1) and guide RNA expression elements.

Fig. 2

a Genomic structure of the uchl1 gene and selection of the gRNA targeting site. To assess our strategy, we performed gRNA/Cas9-mediated knockout of the uchl1 gene on the locus chr4:41,256,413–41,268,455 (GRCh38/hg38), size: 12,043 bases, plus-strand orientation (NCBI Ref: NC_000004.12). b Design of the sgRNA/Cas9 expression construct. To construct the UCHL-1 gRNA/Cas9 expression vector, we used the PX459v2.0 vector (Addgene cat.# 108292). A 5′-adjacent region of exon 3 was targeted and is shown as blue letters in the sequence. The guide RNA sequence is underlined, and the PAM sequence (3 bp NGG) is indicated in the black box. The annealed gRNA oligos were subcloned between two BbsI sites downstream of the U6 promoter.