Figure 1
From: Internal epitope tagging informed by relative lack of sequence conservation
Target site selection for epitope tagging and mutagenesis.
(a) Alignment of the zebrafish, fugu, Xenopus, chicken and human Tcf21 protein sequences. Amino acids identical between all species are in red, identical between at least three out of five species in blue. Absent amino acids (length variation) are highlighted in yellow. Selected target for tag integration is shown by a blue triangle. (b) Alignment of the zebrafish, fugu, Xenopus, chicken and human Tbx18 protein sequences. (c,d) DNA sequences of tcf21 (c) tbx18 (d) target sites (top) and repair oligonucleotides (bottom). sgRNA target sequence is highlighted in cyan, PAM sequence is shown in bold, location of the Cas9-induced DSB is shown by a red line, restriction enzyme site is underlined, homology arms are in magenta, and sequence of the V5 tag is in blue.
