Figure 1
Generation of Tshr mutant rats by CRISPR/Cas9 technology. (A) The diagram showed the TSHR protein domains, including extracellular domain (ECD), transmembrane domains (TMD), and one intracellular carboxy terminal. The gray line refers to the ECDs encoded by exon 1–9, while the black line refers to the domains encoded by exon 10. The scissor indicates the gRNA targeting sequence near the end of 1st intracellular loop (ICL). (B) DNA sequences of the gRNA targeting sites. The sgRNA targeting sequence is shown in black, and the PAM sequence is shown in blue. (C) The targeting and cutting efficiency of CRISPR was tested by a T7 endonuclease I (T7EI) assay in vitro. The image was cropped from different parts of the same gel. The full-length gel is presented in Supplementary Fig. 3. (D) Sequencing results of the mutant alleles in the founders. the PAM squence is highlighted in blue. The deletions are indicated as “−”, whereas the insertions are highlighted in red. The number of deleted nucleotides are shown to the right of each allele.
