Targeted insertions achieved by co-injection of single-stranded oligonucleotides (ssODNs) and the RGN system.

<p>The sgRNAs targeting <i>fh</i> and <i>gsk3b</i> have been described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068708#pone.0068708-Hwang1" target="_blank">[11]</a>. For each gene, the wild-type sequence is shown at the top with the target site highlighted in yellow and the PAM sequence highlighted as red underlined text. For some cases the target sites are highlighted in green if the target sequences are in the reverse complement strand. The ssODNs containing 3–4 nucleotide (nt) insertions are shown beneath the wild-type sequences. The targeted insertions are highlighted as blue underlined capital letters. The target gene sequences identified in the injected embryos are shown beneath the ssODN sequences. Some of them contain only the precise intended changes (labeled as “precise” in parentheses on the right), while others contain additional indel mutations (deletions are shown as red dashes highlighted in grey and insertions as lower case letters highlighted in blue). The number of times each mutant sequence was isolated is shown in brackets.