Figure 1.

Features of assay for measuring gene editing. (A) Sequence of eBFP and eBFP converted into a new GFP (nGFP) coding Tyr⁶⁶ and Phe¹⁴⁵. eGFP sequence is also indicated to show the difference from eBFP and nGFP at codon 145. (B) Schematic representation of ONs tested for targeted mutagenesis. Nomenclature: as, antisense; s, sense; ds, double-stranded; PO, phosphodiester; PS, six phosphorothioate linkages starting from 5′ and 3′ ends; asterisk, phosphorothioate linkage; L, LNA nucleotide; LNA, LNA nucleotide at 5′ and 3′ end position; 1x0-LNA, LNA nucleotide at 5′ end only; 0x1-LNA, LNA at 3′ end only; [1x1]-LNA, LNA nucleotides at internal nucleotide positions 5 and 20; U, three 2′-O-methyl uracyl RNA base starting from 5′ and 3′ ends; branched-PS-eGFP, ON with specific structure consisting of double-stranded DNA with 5′ single-stranded DNA tail coupled together by a baseless deoxyribose branching monomer described as B7 ON in (5). 430-ds-eGFP: 1–430 bp of eGFP sequence. Sequences were as follows: 25-eGFPas, 5′-ACTGCACGCCGTAGGTCAGGGTGGT-3′; 25-eGFP-s, 5′-ACCACCCTGACCTACGGCGTGCAGT-3′; 45-eGFPas, 5′-CGGCTGAAGCACTGCACGCCGTAGGTCAGGGTGGTCACGAGGGTG-3′; 45-eGFPs, 5′-CACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCG-3′; 25-PDEas, 5′-ACTTTCTGCTACGTAGGTTGGAAGG-3′; 25-PDEs, 5′-CCTTCCAACCTACGTAGCAGAAAGT-3′.