Figure 1.

Generation of the LHX6 reporter lines. (A) Schematic overview of LHX6 targeting strategy. The darker green and pale green bars indicate coding parts of exons and 3′ UTR, respectively. The sequence underlined denotes the three gRNAs, while the stop codons are shown in red. The narrower green bars in the targeted allele represent part of the exon or 3′ UTR outside the homology. The arrowed lines under ‘targeted allele’ indicate the positions and predicted size of PCR amplicon of the 5′ and 3′ primer pairs. (B) Genomic PCR detection of the WT and targeted allele using the 5′ primer pairs. (C) PCR verification of Neo cassette removal. (D) An example of sequence read of 5′ PCR product in a targeted clone confirming in frame integration of p2A-mEmerald immediately downstream of the last coding codon of LHX6.