Figure 1. GHRH knockout with CRISPR technology. Location of guide RNAs with respect to exon 2 and exon 3 of GHRH and DNA sequencing chromatogram of mutant GHRH gene between exon 2 and intron 3. (A) Identification of mutations introduced by CRISPR/Cas9 in GHRH gene in founder animals by PCR analysis. (B) 10 G0 pups were tested for indels or deletions. 28528 had a 291 base pairs deletion that eliminates the splice donor site at exon 2, intron 2-3 and a large part of Exon 3 (77 base pairs out of 102 base pairs), showed successful germline transmission. (B) Body weights of female (C) and male (D) WT and GHRH-/- mice from weaning to adulthood. Food intake per mice per day of female (E) and male (F) WT and GHRH-/- mice. Female WT n=11, GHRH-/- n=14, male WT n=11, GHRH-/- n=15. Each bar represents mean ± SEM. Statistical analysis was performed by unpaired Student’s t-test with Welch’s correction; ****p<0.0001.