Fig. 3

RNA-guided CRISPR/Cas9 to knock out miR-274 in PSG. a Design of gRNA. The gRNA site and Drosha site are shown in red arrows. b Schematic diagram of transgenic 2gRNA overexpression vector. c Screening of positive miR-274-2gRNA at embryo stage. (a) Observed under blue light; (b) Observed under white light. d Screening of positive miR-274-2gRNA at adult stage. (a) Observed under blue light; (b) Observed under white light. e Screening of positive miR-274 knockout individuals. (a) Screening of positive miR-274-2gRNA under blue light at embryo stage; (b) Screening of positive Cas9 under green light at embryo stage. (c) Positive miR-274 knockout individuals. f Expression of miR-274 in the PSG of knockout strain. g Base deletion at the site of gRNA1. h Base deletion at the site of gRNA2